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BioNews 35 ENGLISH

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1. eppendorf In touch with life The DIN A2 Poster Maintenance of Centrifuges Order no AQ18636020 is an ideal complement to your manual If you wish to decorate your lab wall with it please ask your Eppendorf sales rep for a personal copy Fixed angle rotors free of charge e Load symmetrically and balance weights BioNews No 35 Innovation KAY KORNER EPPENDORF AG 384 White Wells for Your Real Time PCR The new Eppendorf twin tec 384 real time PCR plates combine all the advan tages of the normal twin tec plates with 384 white wells for your real time PCR New Eppendorf twin tec 384 real time PCR plates The white wells reflect fluorescence much better than clear wells enabling the measurement of samples with lower fluorescence intensity In each single well White wells also greatly reduce interfering background fluorescence leading to improved homogeneity of replicates and increased reproducibility of results New Masterclear real time PCR film New Masterclear real time PCR Film We recommend Masterclear real time PCR Film adhesive as the ideal sealing option for your real time PCR plates e High light transmission gt 90 be tween 450 nm and 750 nm for sensitive real time PCR e Effective adhesive sealing prevents evaporation during real time PCR e Residue free removal from the plate e PCR clean certified free of human DNA DNase RNase an
2. enriching the portfolio with a broad spectrum of innovative equipment for culture growth detection and storage Known for dependable operation year after year New Brunswick s shaker CO2 incubators ultra low temperature freezers fermentors and bioreactors are used in a wide range of in research and commercial applications New Brunswick Scientific s 2011 catalog is packed with innovative new equipment in virtually every product line You can order it by using the fax form on page 15 Shakers CO Incubators Freezers Fermentors Bioreactors and more GN New Brunswick oped cpg Installation and Operational Qualification Installation Qualification IQ and Operational Qualification OQ ser vices assure that your Eppendorf instrumentation is delivered installed and running according to manufac turer specifications With the Installa tion Qualification we document that all items have been delivered and installed as specified Operational Qualification uses the manufacturer s specified procedures and equipment for testing process critical measured values to ensure that your instruments are in a safe and optimal operating condition Eppendorf certification documents include relevant data conforming to Government and other regulatory internal laboratory requirements FDA ISO GxP SOP etc They are supplied to help you prepare for your quality and regulatory docum
3. 280 nm following incubation in 1 5 mL tubes at 95 C for 30 min Values displayed represent the theoretical amount of dsDNA ug mL calculated from the extinction values The resulting LOD limit of detection was determined at 0 006 E 0 3 ug mL the LOQ limit of quantification was determined at 0 021 E 1 0 ug mL Extinction of water following incubation at 95 C for 30 min 0 060 E Eppendorf E Manufacturer V 0 050 2 Or Ot Manufacturer A1 E Manufacturer A2 c 0 040 Q E 0 030 Lu LOQ 260 nm 0 020 0 010 LOD 260 nm 0 000 A 260 A 280 Wavelength nm Application Support Page 2 Europe International Tel 49 1803 666 789 0 09 min aus dem dt Festnetz Mobilfunk max 0 42 min E Mail support eppendorf com North America Tel 1 800 645 3050 E Mail techserv eppendorf com Samples treated in Eppendorf Tubes did not show significantly compromised values Here as in a number of published investigations based on sensitive cell based assays differences between plastic consumables by different suppliers became evident By omitting lubricants and other additives during production Eppendorf tubes are very well suited for sensitive detection methods in the labo ratory Literature 1 McDonald GR Hudson AL Dunn SM You H Baker GB Whittal RM Martin JW Jha A Edmondson DE Holt A Bioactive contaminants leach from disp
4. Canadian scientist Dr Christopher Gregg Postdoctoral Fellow at Harvard University on win ning the 2010 Eppendorf amp Science Prize for Neurobiology for his research on maternal and paternal gene expression in the brain Dr Gregg s work focuses on genes that alter their expression in the brains of offspring according to whether they were inherited from the father versus the mother Understanding the nature of parental effects on gene expression is potentially important for uncov ering the basis of complex human neurological diseases such as autism and schizophrenia as well as eating disorders eppendorf The annual international US 25 000 amp Science Eppendorf amp Science Prize for Neu PRIZE FOR NEUROBIOLOGY robiology honors young scientists for their outstanding contributions to neurobiological research Dr Gregg is the ninth recipient of this prestigious award He was honored at a ceremony held on 15 November 2010 in San Diego USA More information at www eppendorf com prize Applications Rewarded Got a great application Do you have an innovative application enhanced by Eppendorf products Then tell us about it If your application is found to be suitable we will do our best to have it pub lished either on our website in an upcoming issue of Eppendorf BioNews or in a scientific journal Get a great reward In case of publication you will get 100 ep points as well as free Eppendorf products wo
5. Microbiol Infect 14 3 14 2008 8 Schmitz RPH et al New approaches in fungal DNA preparation from whole blood and subsequent pathogen detection via multiplex PCR In Y Gherbawy K Voigt eds Molecular identification of fungi Springer Verlag pp 317 335 ISBN 978 3 642 05041 1 2009 9 Bruns T et al Identification of bacterial DNA in neutrocytic and non neutrocytic cirrhotic ascites by means of a multiplex polymerase chain reaction Liver Int 29 1206 1214 2009 10 Schmitz RPH Lehmann M Pan bacterial detection of sepsis causative bacteria In Molecular detection of human pathogens D Liu ed Taylor amp Francis CRC Press in press 2010 11 Further details can be taken from the Eppendorf Application Note 231 at www eppendorf com applications Readers service Mastercycler pro Ref no 224 BN 35 July 2011 Asia Pacific Tel 60 3 8023 6869 E Mail support_asiapacific eppendorf com Eppendorf BioNews Application Notes Automated Plant DNA Purification Using the Nucleon Plant DNA Kit on the epMotion 5075 TMX Alison Hobday Botwright and Sheila Doyle Gen Probe Life Sciences Ltd Manchester United Kingdom Eric Cairns and Warren Higgins Gen Probe Life Sciences Ltd West Lothian United Kingdom Introduction In the field of plant molecular biology successful analysis requires the purifica tion of reliable and high quality genomic DNA The purification of such DNA can be time consumi
6. Micromanipulation set up PatchMan NP 2 Eppendorf CellTram vario Eppendorf CustomTip Type l 10 um blunt end capillaries Eppendorf CellSurgeon Rowiak Hanover Germany equipped with an Inverted microscope AxioObserver D1 Zeiss Jena Germany T pulse 200 10 MHz 2 5 W 1030 nm Amplitude Systems Pessac France Camera Jenoptik Jena Germany Animals Medaka embryos line 2216 with m cherry red labeled sensory neuro secretory cells K Tessmar Raible Other material 35 mm Petri dish with 0 17 mm glas bottom WillCo Wells Amsterdam Netherlands Application Support Europe International Tel 49 1803 666 789 0 09 min aus dem dt Festnetz Mobilfunk max 0 42 min E Mail support eppendorf com North America Tel 1 800 645 3050 E Mail techserv eppendorf com 6 methyl cellulose Embryo rearing medium 1x ERM 4 Tricaine methanesulfonate stock solution Preparation of 35 mm dishes and adjust ment of micromanipulation set up In order to access the forebrain in the flattest angle possible dishes were cut down to a height of 4 to 5 mm and the capillary holder was inserted in the lower mount on the X head The angle for the universal capillary holder was adjusted in such a way that is was pos sible to move the capillary directly along the glass bottom until it reached the embedded embryo on the same focal level This was necessary in order to avoid shifting the position of the embr
7. Page 8 Spectrophotometric assessment of cross contamination indicates that no DNA can be detected from the 48 cabbage leaf sample preparations in the negative control wells Table 2 This finding was further supported by gel analysis Fig 3 Fig 3 Evaluation of cross contamination by agarose gel electrophoresis of purified DNA samples from cabbage leaves processed in alternative wells The wells AM CM EM and GM are the negative buffer control BM DM FM and HM are the Lambda DNA positive control RNA contamination is evident in some gels This may be avoided with a longer incubation with RNase A and bead mixture or increased levels of RNase A Application Support Europe International Tel 49 1803 666 789 0 09 min aus dem dt Festnetz Mobilfunk max 0 42 min E Mail support eppendorf com North America Tel 1 800 645 3050 E Mail techserv eppendorf com The integration of the Nucleon Plant DNA Kit with the epMotion 5075 TMX platform provides a reliable convenient and flexible system for the automated purification of high quality high yield DNA from up to 96 plant samples in approximately 4 hours with minimal hands on time This easy to handle system benefits not only from increased throughput but also reproducible purification of high quality high molecular weight DNA for use in sensitive downstream applications Literature Application Note 230 Automated Plant DNA Purification using the N
8. a Set of 3 pipettes when you participate in our competition We hope you enjoy the read Your BioNews editorial team carbon neutral Gh natureOffice com DE 299 920637 print production 2 BioNews No 35 IMPRINT Editorial team Berrit Hoff Editor in Chief Axel Jahns Jochen Muller Ibeler Natascha WeiB Publisher Eppendorf AG Barkhausenweg 1 22339 Hamburg Germany Telephone 49 40 53801 636 Fax 49 40 53801 840 E Mail bionews eppendorf de Internet www eppendorf com We welcome all readers articles for this publication However no responsibility is accepted for unsolicited manuscripts Technical specifications subject to change Errors and omissions excepted Important note The new products described may be launched at different times in various countries Please contact your local distributor for details Copyright All rights reserved including graphics and images Copyright by Eppendorf AG July 2011 Contents Cortifuge S424 A In the spotlight Innovation Straight from the lab News Tips Service I 1 Natascha WeiB Wolf Wente Stefanie Topp I emacs 1 The Influence of UV Absorbing Substances Released I I I The 1 tie of a UV Abso ane rbing Substan iners Le ati ables on AEn a Raed om Pasti from Plastic Containers Leachables on Photometric Analyses 1 2 Kristin Tessmar Raible Katharina Sc
9. certainty they need in addition to their own experiences This includes instru 7 Ze User friendly TUV e User tested gt TOV Rheinland is i 974209 ments that users can consistently depend on because they know them inside out classics like the Reference Pipette Dependable safe ergonomic The Reference s continuous success is largely due to its unique design and technology concept With its control button and ejector combination the pipette is designed to actively reduce contamination from aerosols All this is complemented by a high level of ergo nomics the pipette has only one control button and the volume can be easily read regardless of the position of the hand The Reference not only meets the high criteria of the Eppendorf PhysioCare Concept its user friendliness has also been certified by the renowned TUV Rheinland Germany Positive feedback from countless users confirms The Eppendorf Reference continues to be the reference class for modern liquid handling tools With the utmost precision and accuracy low failure rate and high longevity it continues to please old and new fans maintaining its strong position in the Eppendorf pipette portfolio The Reference on the internet www eppendorf com reference Eppendorf Reference e Ref no 30 EED BioNews No 35 ie Nano ls All Around Nanotechnology and Biotechnology explained clearly Have you ever wo
10. of pathogen specific targets Standard multiplex PCR was performed within two pools of specific primers The Application Support Europe International Tel 49 1803 666 789 0 09 min aus dem dt Festnetz Mobilfunk max 0 42 min E Mail support eppendorf com North America Tel 1 800 645 3050 E Mail techserv eppendorf com assay covers 34 bacterial and 6 fungal species including Bacteroides fragilis Burkholderia cepacia Aspergillus fumi gatus Candida albicans Clostridium perfringens Enterococcus faecalis Enterococcus faecium Escherichia coli Haemophilus influenzae Klebsiella oxytoca Neisseria meningitidis Proteus mirabilis Pseudomonas aeruginosa Serratia marcescens Staphylococcus aureus Staphylococcus epidermidis Staphylococcus haemolyticus Stenotro phomonas maltophilia Streptococcus agalactiae Streptococcus mutans Strep tococcus pneumoniae Streptococcus pyogenes Streptococcus sanguinis 11 In addition a set of frequent antibiotic resistances e g methicillin vancomycin and 8 lactamases are also targeted specifically Bacillus subtilis specific primers are within each pool as internal run control The PCR was set up using the VYOO kit PCR reaction vessels which contain the lyophilized primer mix In addition a PCR reaction contained 1x QIAGEN Multiplex PCR Master Mix QIAGEN Hilden Germany and 1 ug pathogen enriched DNA filled up with DNA DNase free water No template contro
11. pipette tips have not been modified and the new epT I P S Motion are avail able as in the past with and without filter and in different purity grades All details about the new epT I P S Motion box system are available at www epmotion com or in the brochure which you can re quest using the reference number de noted below epT I P S Motion e Ref no 180 epT I P S Motion Rack color coded tray in a new box epT P S Motion Reload refill system composed of Reload tray above and reusable TipHolder Adapter below epMotion Performance Plans Our Performance Plans for epMotion are maintenance programs that are tailored to address the different levels of demand They include cleaning inspection partial overhaul calibration and adjustment according to the speci fications Calibration of the dispensing tools can be performed according to ISO 8655 with NIST traceable standards supporting GLP documentation Furthermore we offer certification services for regulated areas including Installation and Operational Qualifica tion IQ OQ documentation as well as certified training and application support The service work and training application support is performed by our professionally trained Service Technicians and Application Specialists ep Performance Plans features e PREMIUM Plans for two visits year includes 1 year warranty extension e No hidden fees makes c
12. 95 C for 30 min Application Support Europe International Tel 49 1803 666 789 0 09 min aus dem dt Festnetz Mobilfunk max 0 42 min E Mail support eppendorf com North America Tel 1 800 645 3050 E Mail techserv eppendorf com Asia Pacific Tel 60 3 8023 6869 E Mail support_asiapacific eppendorf com Page 1 Eppendorf BioNews Application Notes The Influence of UV Absorbing Substances Released from Plastic Containers Leachables on Photometric Analyses Comparable results were obtained for the PCR tubes the water taken from competitors tubes treated in the thermo cycler yielded higher extinction values than the water incubated at room tem perature Fig 1b According to these data it appears that substances which absorb light in the UV range are released from the com petitors containers tested herein The maximum values are observed slightly above 190 nm In addition water from the 1 5 mL tubes showed absorption signals in the range of 210 220 nm as well as 260 nm Fig 1a It is exactly this set of wavelengths which plays an important role during photometric detection and quantification of biomolecules For example proteins may be measured directly at 205 nm or at 280 nm whereas nucleic acids are measured at 260 nm One consequence of leached UV absorbing components is illustrated in Fig 2 Here extinction values of water incubated at 95 C ina further experiment are shown at 260 nm
13. ININ WVGETAEDINTAN Mean consistently high purity DNA with a Sample calles Paks eae Eo oa S mean 260 280 absorbance ratio of 2 05 indicating low protein contamina anode oe BG ed eae S ee tion in the resulting DNA samples The average total yield across the Empty j a He eee z sample DE derived from 30 mg of Table 2 Spectrophotometric assessment of cross contamination starting plant material was found to be 11 65 ug Cross contamination Conclusion Quality of DNA and structural integrity In order to further demonstrate quality and structural integrity of the isolated DNA the purified DNA samples were analysed by agarose gel electrophoresis Fig 1 A7 A8 A9 A10 A11 A12 Fig 1 Agarose gel electrophoresis of purified DNA samples from cabbage leaves A1 A6 pine needles A7 A12 rose leaves B1 B6 and corn B7 B12 negative buffer control Lambda DNA positive control The cabbage sample was further ana lysed with and without treatment with restriction enzyme The automated purification procedure was shown to produce high molecular weight DNA as indicated by the clear bands Fig 2A Efficient restriction enzyme digestion was achieved with the gels producing the characteristic smear of DNA Fig 2B Fig 2 Gel electrophoresis of purified DNA samples from cabbage leaf without A and following restriction digestion B Lanes 1 4 purified cabbage DNA Lane 5 negative control Lane 6 DNA ladder
14. Products News BERRIT HOFF EPPENDORF AG Learning with Eppendorf Products DNA Learning Centers in the USA For more than twenty years Eppendorf has been supporting the Dolan DNA Learning Center DNALC the world s first science center devoted entirely to genetics education and an operating unit of Cold Spring Harbor Laboratory located on Long Island New York This support continues through Eppendorf North America ENA as the DNALC adds its third and youngest facility designed to serve Manhattans inner city in Harlem NY USA The Harlem DNA Lab was equipped with a wide range of Eppendorf laboratory products This included almost 100 pipettes four microcentrifuges several cyclers and a variety of pipette tips and Eppendorf tubes reports Dan Decker Vice President of Sales for ENA and a member of the DNALC corporate advisory board The Harlem DNA Lab is a cooperation of the New York City Department of Education and the DOLAN DNA Learning Center Cold Spring Harbor Laboratory In the newly created laboratory school groups from New York City and the surrounding areas cover broad topics within the field of genetics An extensive continuing education program is also offered for teachers Since the founding of the DNA Learning Centers in 1988 more than 325 000 students and 8 000 teachers have profited from their program naturally using Eppendorf products DNA Visitors Laboratory at Deutsches Museum Mun
15. _asiapacific eppendorf com clusters in order to excise those in cubi cal fashion Excised cell clusters were made up of an estimated 75 of cells of interest so that further molecular biological analysis such as transcrip tome analysis would be possible The high resolution of microsteps offered by the PatchMan NP 2 allowed precise positioning of the microcapillary before as well as after excision Successful removal of cell clusters was feasible as CellTram vario offers a very small minimum adjustment volume fine drive and therefore allows to generate a vacuum just right to remove cells but without detaching single cells from clusters The above described method could be of great value to researchers working with transgenic small model organisms trying to unravel genetic traits Literature 1 Wittbrodt J et al 2002 Medaka a model organism from the Far East Nature Reviews Genetics 3 53 64 Readers service PatchMan e Ref no 52 CellTram Ref no 77 BN 35 July 2011 Eppendorf BioNews Application Notes Rapid and Reliable Multiplex PCR Pathogen Detection in Whole Blood by Using the VYOO Kit on the Mastercycler pro Kristin Wessel and Roland P H Schmitz SIRS Lab GmbH 07745 Jena Germany Abstract Here we introduce a new protocol for multiplex PCR based detection and identification VYOO SIRS Lab Jena Germany of bacterial and fungal patho gens directly from whole blood As s
16. am vario and Eppendorf PatchMan NP 2 Introduction Medaka is a well established model organism for the analysis of cell signal ling in developmental and neurobiologi cal research 1 Transgenic animals expressing fluores cent proteins under specific enhancer promoters are of special interest as they provide a tool to obtain a better under standing of control mechanisms during different stages of development In this context the analysis of the impact of environmental factors on the cell s tran scriptome is of great interest Such transcriptome analyses can be per formed by modern technologies includ ing next generation sequencing In order to be successful starting material that is July 2011 BN 35 2Rowiak GmbH Hanover Germany as pure as possible is required How ever gaining cell material of such quality from small pieces of tissue such as a Medaka forebrain is a challenging task We approached this challenge by using Rowiak CellSurgeon in combination with an Eppendorf micromanipulator set up Rowiak CellSurgeon is a laser scanning microscope with cutting ability which was used to image measure and excise cell clusters of sensory neurosecretory cells expressing m cherry red from the forebrain of Medaka embryos Excised cell clusters were then removed with a 10 um microcapillary controlled by PatchMan NP 2 micromanipulator and CellTram vario microinjector Material and methods
17. and 280 nm In addition the theoretical amount of dsDNA which would result from this value is shown The value for the samples obtained from the Eppendorf Safe Lock Tubes was below the detec tion level LOD limit of detection whereas the water from the other tested vessels yielded values above the limit of Absorbance scan of water following cycling and incubation at RT 2 25 0 30 2 00 4 0 25 1 75 soe 1 50 0 15 QO Q as 0 10 25 7 g 0 05 2 1 00 _ 0 00 E in 240 260 280 300 0 75 I 190 210 230 250 270 Eppendorf RT Eppendorf Cycler Manufacturer T RT Manufacturer T Cycler Manufacturer A RT Manufacturer A Cycler 290 310 330 350 370 390 Wavelength nm Fig 1b Absorbance spectrum of water following incubation of 0 2 mL PCR tubes in the cycler and at room temperature RT quantification LOQ Therefore elevated results for DNA concentration would be obtained during DNA quantification When the actual amount of DNA present is lower than the measured value a neg ative impact on downstream applications may ensue Conclusion Photometric analyses verified that UV absorbing substances are released from plastic vessels made by certain manufacturers and that they interfere with the detection of biomolecules such as nucleic acids and proteins Fig 2 Extinction of water at 260 nm and
18. apid tool for detection of low level infections It supports and expedites clinical findings and strengthens a directed antibiotic therapy Application Support Europe International Tel 49 1803 666 789 0 09 min aus dem dt Festnetz Mobilfunk max 0 42 min E Mail support eppendorf com North America Tel 1 800 645 3050 E Mail techserv eppendorf com Literature 1 Reinhart K et a Diagnosis and therapy of sepsis Clin Res Cardiol 95 429 454 2006 2 Ibrahim EH et al The influence of inadequate antimicrobial treatment of bloodstream infec tions on patient outcomes in the ICU setting Chest 118 146 155 2000 3 Fine JM et a Patient and hospital charac teristics associated with recommended proc esses of care for elderly patients hospitalized with pneumonia Arch Intern Med 162 827 833 2002 4 Garnacho Montero J et al Impact of ad equate empirical antibiotic therapy of patients admitted to the intensive care unit with sepsis Crit Care Med 31 2742 2751 2003 5 Valles J et al Community acquired blood stream infection in critically ill adult patients impact of shock and inappropriate antibiotic therapy on survival Chest 123 1615 1624 2003 6 Iregui M et al Clinical importance of delays in the initiation of appropriate antibiotic treat ment for ventilator associated pneumonia Chest 122 262 268 2002 7 Chastre J et al Evolving problems with resistant pathogens Clin
19. as position 1 using the PatchMan NP 2 The microcapillary was withdrawn a few millimeters and then slightly moved upwards Laser assisted excision of cell clusters Cell clusters were imaged in laser scanning mode at 40x magnification to determine their x y and z dimensions The parameters established were then used to excise cell clusters in cubical fashion and concomitantly to minimize Fig 2 Page 4 contamination with unlabeled cells by choosing cutting lines as close as pos sible to cell clusters See Fig 2 Removal of cell clusters As shown in Fig 3 excised cell clusters were imaged in fluorescence mode at 20x or 40x magnification and the micro capillary was slowly fine or extra fine mode brought back to position 1 Cell clusters were aspirated using the fine drive of the CellTram vario The microcapillary was slowly withdrawn and then moved to the Home position Cell clusters were transferred into a microcentrifuge tube turning the dial for fine drive clockwise Results and discussion With the use of laser scanning micros copy Rowiak Cell Surgeon we were able to determine dimensions of cell Fig 3 Application Support Europe International Tel 49 1803 666 789 0 09 min aus dem dt Festnetz Mobilfunk max 0 42 min E Mail support eppendorf com North America Tel 1 800 645 3050 E Mail techserv eppendorf com Asia Pacific Tel 60 3 8023 6869 E Mail support
20. d PCR inhibitors More information Learn all there is to know about Eppendorf PCR plates at www eppendorf com pcrplates or request the latest brochure on real time PCR consumables The bright choice using the reference number denoted below The bright choice brochure Ref no 223 8 BioNews No 35 Interference with DNA Measure ments by Plastic Consumables It is nearly impossible to imagine our daily life without synthetic materials In molecular and biochemical labora tories consumables made from plastic are used in practically every application However in the recent past scientific reports of skewed DNA measurement results due to certain leachable additives which are used during production by some manufacturers of plastic consumables have accu mulated Not only sensitive enzymatic assays are affected As described in the Application Note on p 1 2 simple heating of the sample in plastic containers made by certain manufacturers can lead to leaching of substances which critically interfere with the detection of biomolecules such as DNA and proteins and which may thus skew concentration measurements Warming of samples occurs during many routine applications in molecu lar or cell biology such as cell lysis centrifugation or PCR By avoiding slip agents biocides plasticizers and other substances which may interfere with DNA measurements Eppendorf tubes and tips are id
21. ded For the purpose of this Technical Report experiments were performed in accord ance with the publication by Lewis et al 3 This publication had described that the Eppendorf vessels tested achieved noticeably better results than competi tors but no data were shown Hence Eppendorf Safe Lock Tubes 1 5 mL and Eppendorf 0 2 mL PCR tubes were here tested alongside comparable containers from other manufacturers Following incubation of water at different tempera tures absorbance scans and measure ments in the UV range were performed in order to test whether components were leached which could compromise subsequent photometric analyses July 2011 BN 35 Extinction can be estimated to be pro portional to the amount of leached sub stances Materials and methods Three Eppendorf Safe Lock Tubes 1 5 mL and comparable tubes from two competitors were filled with 1 mL water molecular biology grade and incubated under the following conditions a 95 C for 30 min b 70 C for 30 min c 37 C for 24 h 0 2 mL PCR tubes from Eppendorf and two other manufacturers were filled with 150 uL water molecular biology grade each One half of the containers was subjected to a PCR protocol in a thermo cycler 30 PCR cycles 95 C 30 s 60 C 30 s 72 C 2 min while the remaining vessels were incubated at room temper ature RT Each sample was subjected to ascan across the range of wave lengths between 190 nm a
22. duced by 25 compared to the previous model The centrifuge saves up to 60 in power consumption with the FastTemp function for quick pre cooling epGreen Would you like additional information The Centrifuge 5424 R brochure can be ordered from us using the ref no denoted below Detailed information is also available at www eppendorf com centrifugation Centrifuge 5424 R e Ref no 235 BioNews No 35 GC Straight from the lab Safe and Efficient Centrifugation JAN HENDRIK BEBERMEIER EPPENDORF AG Safe and Efficient Centrifugation Do you already know the new Eppendorf poster Main tenance of Centrifuges It provides you with useful advice on the proper cleaning and maintenance of your Eppendorf centrifuge as well as instructions for the correct rotor loading and other interesting and smart MINN A I tips for safe and efficient centrifugation Here are some examples from the contents Fasten rotor tightly Prior to centrifugation rotor must be tightened securely on drive shaft using a rotor key For swing bucket rotors ensure that buckets are properly hooked onto the rotor Perform a manual swing out test to check that the buckets are moving freely In swing bucket rotors buckets with the same weight class must be on opposite positions To check the weight category of the bucket check the value at the side of the buckets Note the weight specifica a tions printed on the
23. eally suited for all sensitive detection methods in the laboratory www eppendorf com consumables Therefore rely on Eppendorf quality for reproducible and reliable results DANIEL WEHRHAHN EPPENDORF AG Eppendorf BioNews Application Notes The Influence of UV Absorbing Substances Released from Plastic Containers Leachables on Photometric Analyses Natascha Wei Eppendorf AG Hamburg Germany Wolf Wente and Stefanie Topp Eppendorf Instrumente GmbH Hamburg Germany Introduction A 2008 Science publication and other papers described that slip agents as well as biocides which were washed from vessels and tips are able to show activity in specific enzyme assays 1 2 As a consequence false positive or false negative results are produced which cannot be analyzed thus leading to increased consumption of time and financial resources Even routine applications have been shown to be compromised 3 UV absorbing substances from plastic containers are leached into the sample by laboratory applications requiring temperatures of 37 C or above Since these substances absorb light in the same range as the absorbance maxima of nucleic acids and proteins they can interfere with photometric detection reactions thus providing a source of error with adverse effects on down stream applications Therefore the use of high quality con sumables which contain the least amount of leachable additives is recom men
24. emains constant even at a maximum speed of up to 21 130 x g or Centrifuge 5424 R 15 000 1 min al BioNews No 35 Silence Speed Simplicity Centrifuge 5424 R In the spotlight Awarded annually since 1955 the red dot award recognizes global design achievements for a wide variety of in dustrial products Silence I Speed I Simplicity The Centrifuge 5424 R doesn t just stand out with its excellent ergonomics it perfectly embodies our motto Silence Speed Simplicity It is even quieter faster and easier to operate than its predecessor Silence Quiet for a comfortable working environment All mechanical components have been carefully selected and optimally com bined to create a device with significantly reduced operating noise The result the Centrifuge 5424 R is incredibly quiet even during operation without rotor lid Centrifuge 5424 R with keypad The rotor chamber s new OptiBowl design markedly reduces background noise to ensure a pleasant working environment Speed Faster and more flexible for all applications The powerful Centrifuge 5424 R offers a high centrifugation speed of up to 21 130 x g 15 000 1 min in a tempera ture range of 10 C to 40 C Quick precooling e g from 21 C to 4 C can be achieved in just 8 minutes with the FastTemp function Moreover a secure SOFT brake provides maximum protec tion of sensitive samples Simplicity Safety via intui
25. entation and audits Your value e Ensures the safety and quality of your work e Proof of compliance with the rel evant technical specifications e Support for your regulatory tasks and GxP requirements ep Contact your local Eppendorf office for more information or visit www eppendorf com epservices and or your local Eppendorf website BioNews No 35 an Service Prize Competition Win online at www eppendorf com bn service Prize Competition The solution of the prize competition of BioNews No 33 was Eppendorf LoBind Claudia Braunig f2mb GmbH Bad Lan gensalza Germany won the first prize an Eppendorf Xplorer pipette You can either use the reply fax p 15 send us an e mail to bionews eppendorf de or participate online at www eppendorf com bn service All correct answers will be considered for a prize Winners will be notified in writing Cash payment of the prize is not possible No recourse to legal action The judges decision is final Eppendorf employees and their families may not participate The winner of the first prize will be published in BioNews No 37 Have fun in our new crossword How to find out the solution Simply arrange the encircled letters of the crossword in the correct order Send us the solution until 31st October 2011 1st prize 1 epReference Pack including 3 Eppendorf Reference pipettes 0 5 10uL 10 100uL 100 1 000 uL matching epT I P S 3 pi
26. eppendorf l EAT UAIT UALL FT EET Le EE T The Influence of UV Absorbin I New Box Svstem for epl P S Motion y p i hn caress S I im UV Absorbing Substances Released from Plastic Containers Influencing Photometric Analyses Multiplex PCR Pathogen Detection in Whole Blood Automated Plant DNA Purification etc Editorial Dear Reader Quality reliability experience innovation these are words that people worldwide associate with Eppendorf Whether a new technology a service or the further advancement of an existing product is concerned every detail is geared towards the needs and requirements of our users This approach has been crowned with success in every respect as detailed in the annual report 2010 p 13 The success story of the Eppendorf microcentrifuges continues Just recently our Centrifuge 5424 R was awarded the red dot design award 2011 p 4 5 Our family of consumables has new additions new twin tec Plates with 384 wells for real time PCR as well as epT I P S Motion with a new ecological box system p 8 9 Eppendorf honors young scientists On page 12 we are introducing to your the recent laureates of the Eppendorf research prizes Further to more information and product solutions for a wide range of tasks as always the insert contains detailed Application Notes Last but not least you will have the chance to win
27. epsis is one of the most common causes of death in hospitalized patients a rapid pathogen detection is a corner stone in effective therapy To investigate DNA isolates from whole blood samples of intensive care unit patients for the presence of sepsis causing pathogens a microbial DNA enrichment was performed followed by a standard multiplex PCR on the Mastercycler pro Gel based amplicon analysis confirmed successful nucleic acid trace detection of 34 bacterial and 6 fungal targets as well as 5 antibiotic resistances with an overall sensitivity of 10 to 100 colony forming units cfu mL whole blood Introduction Sepsis results from the host s response to bacterial and fungal infections whereas malfunction of the defense and repair system seems to be responsible for the development of organ dysfunctions In Germany 154 000 patients suffer from severe sepsis each year With 60 000 deaths it is one of the most frequent causes of death in the ICUs About 30 of the intensive medicine budgets are expended for the treatment of those patients 1 A prompt and adequate antibiosis started in the first few hours of infection assigns the crucial step for an effective therapy 2 4 Epidemiological data confirm that a doubling of mortality is the consequence of inadequate ther apies 5 and an increase of mortality of more than 7 per hour is proven in cases of delayed adequate antibiotic treatment 6 In addition developmen
28. hipany Sabine Przemeck Removal of Fluorescently labeled Sensory neurosecretory Cells from Forebrain of Transgenic Medaka Embryos Using Eppendorf PatchMan NP 2 and CellTram vario and Rowiak CellSurgeon 3 4 Ta hn han hs hes hs s s as hes ies ies hes hes ies ien ies en in s s s as s s s s s a a s i s i i a a a Da Da Da Da Da a Da Da Da a s s Da i a es a a 1 Kristin Wessel Roland Schmitz PS SS measur i Rapid and Reliable Multiplex PCR Pathogen Detection in Whole Blood by Using the VYOO Kit on the Mastercycler pro 5 6 Subsequer Alison Hobday Botwright Sheila Doyle Eric Cairns Warren Higgins i i Automated Plant DNA Purification Using the Nucleon Plant DNA Kit on the epMotion 5075 TMX 7 8 i BioNews No 35 E In the spotlight Silence Speed Simplicity Centrifuge 5424 R Silence Speed Simplicity Centrifuge 5424 R The new Eppendorf microcentrifuge standard Centrifuge 5424 R is quieter faster and easier to use than ever True to our motto Silence Speed Simplicity we have developed our new generation of microcentrifuges in line with the strictest requirements for operating comfort and safety And it s paid off launched in 2010 the Centrifuge 5424 R has now been awarded the red dot design award 2011 as the compact 24 place microcentrifuge the successor to the legendary Centrifuge 5415 R offers even more comfort in the laboratory The Ce
29. hoosing and budgeting easy Your benefits e Low risk of operational downtime e High performance in precision and accuracy e Audited system for GLP compliance For more information service requests and local offers go to www eppendorf com epservices and or your local Eppendorf website Performance Plans are available in selected countries only and service offers may differ BioNews No 35 Straight from the lab JANINE JACOBI EPPENDORF AG Reference Class Since 1994 Some things are timeless ever present yet no one thinks about them Like the Eppendorf Reference pipette which has been on the market since 1994 or the Comforpette its predecessor from the 1970s Both are one button pipettes with combined control button and ejector A true companion didn t Know any other pipettes At my position at the Bogenhausen Hospital in Munich only worked with my Com forpette says Kornelia Ewald Respon sible for conducting end user seminars on Liquid Handling at the Eppendorf Training Center Ms Ewald has worked at Eppendorf as Application Specialist since the beginning of the 1990s As one button pipettes continue to enjoy immense popularity they are of course an integral part of the trainings Classic constants Advancements in science research and our everyday lives occur in shorter and shorter cycles It s not easy to keep pace Here true constants give the users the
30. ich Eppendorf also welcomed the opportunity of donating products like pipettes thermomixers and centrifuges to the DNA Visitors Lab at the New Technologies Centre ZNT of the Deutsches Museum in Munich Germany Assisted by young scientists visitors can do their own experi ments in the DNA Visitors Laboratory coming to grips with a pipette and other equipment used in the field of molecular biology That helps them to understand the day to day work of a researcher in the lab while gaining useful knowledge on cell biology heredity and genetic engineering For example par ticipants can use genetic fingerprints to solve a fictitious crime by processing DNA samples from the scene and those of a suspect and then comparing them The program is intended for interested participants in the 9th Class or higher and is also offered as a teacher training session From the course program e Whodunnit Genetic fingerprinting with PCR e Genetically modified Genetic analysis with PCR e Genetic engineering activities inserting a gene into a plasmid SA A _ ee al Understanding genetic research Course participant at DNA Learning Center Harlem New York USA More information at www dnalc org Photo DNALC Harlem Wy n A Course participants in the DNA Visitors Laboratory at Deutsches Museum More informationen at www deutsches museum de en exhibitions new technologies Photo Deutsches Museum B
31. ioNews No 35 g News BERRIT HOFF amp CAROLYN POWELL EPPENDORF AG Eppendorf Honors Young Scientists Eppendorf Award for Young European Investigators 2011 QPORF gt e On 25 May 2011 the Eppendorf Award for Young European Investigators which honors outstanding work in biomedical research in Europe was presented for the 16th time For the first time the award ceremony took place at the EMBL Advanced Training Centre in Heidelberg Germany OA doy gt A Opean Y The 15 000 prize went to Assistant Professor Suzan Rooijakkers University Medical Center Utrecht Depart Medical Microbiology Utrecht The Netherlands for her discoveries of how the pathogen Staphylococcus aureus evades immune attack to survive in the human host She found out that the Staphylococci secrete proteins that block critical steps in the complement cascade One such protein is the unique complement inhibitor SCIN that she found to inhibit the C3 convertase which is required for the complement system to tag the bacteria for destruction These results are creating new inroads into developing drugs against inflammatory and infectious diseases The Eppendorf Young Investigator Award is presented in partnership with Nature More information at www eppendorf com award t WANTED APPLICATIONS _ REWARDED www eppendorf com applications Eppendorf amp Science Prize for Neurobiology 2010 Congratulations to
32. ls NTC were devoid of DNA The PCR reaction was performed on the Mastercycler pro Figure 1 Multiplex PCR amplicons were analyzed on agarose gels and assigned to patho gens and or antibiotic resistances on the basis of pool specific length markers Fi 13 4 ars 4 33 15 Fig 1 Temperature profile of the PCR program on the Mastercycler pro Page 5 Asia Pacific Tel 60 3 8023 6869 E Mail support_asiapacific eppendorf com Eppendorf BioNews Application Notes Rapid and Reliable Multiplex PCR Pathogen Detection in Whole Blood by Using the VYOO Kit on the Mastercycler pro Pool I Pool Il 777 bp 341 bp 285 bp Fig 2 Proof of Enterococcus faecium genomic DNA in EDTA whole blood by VYOO Samples were taken from ICU patients suspected for sepsis M1 2 DNA marker pool I Il 1 sample DNA tested with primer pool l 2 sample DNA tested with primer pool ll 1 Bacillus subtilis specific band at 285 bp for internal run control 2 E faecium specific band at 777 bp vanA resistance specific band at 341 bp B subtilis specific band at 285 bp internal run control Results After preparation of exemplary EDTA whole blood samples from patients sus pected for sepsis agarose gel analysis of PCR products revealed single bands Fig 2 Amplicons were specific for Enterococcus faecium and vancomycin A resistance pool 2 as compared to species specific marker bands Positive co
33. nd 400 nm using a spectrophotometer and the values for the groups of samples were determined Non incubated water was used to set the blank value In an addi tional series of measurements in the BioPhotometer plus 1 mL water molec ular biology grade was incubated at 95 C for 30 min in each of three Eppendorf Safe Lock Tubes and tubes from three competitors Extinction values at 260 nm and 280 nm were determined the mean values of the replicates and the SD were calculated Furthermore the factor 50 ug mL was used to determine the theoretical amounts of dsDNA from the measured extinction values Results and discussion Fig 1 demonstrates that water which was incubated in Eppendorf Tubes shows no significant extinction in relevant range of wavelengths The samples taken from the tubes made by other manufacturers show a distinct temperature dependent absorption profile The highest values were measured at 95 C Fig 1a At 70 C and 37 C these values were lower data not shown Absorbance scan of water following incubation at 95 C for 30 min 2 25 0 10 2 00 T a i 0 08 1 75 c 1 25 1 00 Extinction OD 0 75 0 50 0 25 0 00 I 190 210 230 250 270 290 310 330 350 370 390 Wavelength nm Eppendorf Manufacturer V Manufacturer A1 Fig 1a Absorbance spectrum of water following incubation of 1 5 mL tubes at
34. ndered about the early beginnings of biotechnology How transgenic animals are created Which technology tools are required How two cells become one and how this cell is then used as a workhorse in cancer research These and many more questions are answered by Nano and Biotechnol ogy an impressive permanent exhibit at the Centre for New Technologies of the Deutsches Museum in Munich Germany opened in November 2009 For the explanation of the fascinating possibilities and developments within the life sciences the scientific director of the exhibit Dr Sabine Gerber places high value on realistic presentations which are easy to understand Several areas of the exhibit cover the nanosystem cell and the targeted alteration re programming of these cells in order to employ them in the treatment of disease or the production of pharmaceuticals These methods are demonstrated in part using original Eppendorf cell technology products specifically the Multiporator and a complete micro manipulation work station Both these instruments have become the estab lished standard equipment in scientific laboratories More information about the exhibit can be found at http www deutsches museum de en exhibitions new technologies nanotechnology More information about the topic cell technology may be found at www eppendorf com cell HEIDE NIESALLA EPPENDORF AG Learning with Eppendorf
35. ng particularly in large scale projects where many individual samples need to be processed A reli able reproducible and easy to handle automated method for the purification of plant derived DNA is therefore very desirable In this report we demonstrate an auto mated method for the purification of plant genomic DNA using the Nucleon Plant DNA Kit from Gene Probe Life Sciences on the Eppendorf workstation epMotion 5075 TMX The automated method was found to deliver consistently high purity high molecular weight DNA from the sample types tested With a protocol time of 4 hours with minimal hands on time for 96 samples and no detectable cross sample contamination the methodology provides a viable easy to use automated procedure for the purification of plant derived DNA The Nucleon Plant DNA Kit is designed for the rapid economical purification of high quality high yield DNA from a wide range of plant materials The purification process is based on the reversible bind ing of DNA to novel magnetic beads The beads which are not silica coated have nanometre dimensions which can aggregate to form functional complexes These complexes provide the means to purify DNA from complex mixtures and provide consistent quality and yield The epMotion 5075 TMX includes an integrated TMX module to shake and heat samples which can be loaded and unloaded with the gripper The inte grated TMX module means that following Automation
36. ntrifuge 5424 R was developed Prize winning according to comprehensive ergonomics research to meet users constantly increasing demands Routine tasks can be completed more quickly and easily than ever before For example with the FastTemp precooling function the cen l and ecological requirements trifuge achieves a time savings of 45 the Centrifuge 5424 R has compared to the previous model P p proven its merit with the Latest generation of refrigerated awarding of the red dot microcentrifuges design award 2011 The design and unique ergonomics of the Centrifuge 5424 R have now been honored Subject to the strictest evalua tion criteria which include degree of innovation functionality longevity The versatile compact microcentrifuge 5424 R is designed as a key element of an ideal laboratory working environ ment which guarantees reliable and safe performance according to the highest standards in molecular biology biochemistry and biotechnology With its compact footprint and low access height the Centrifuge 5424 R is ideal for every lab It comes standard with an aerosol tight 24 place rotor for 1 5 2 0 mL tubes and features three additional rotor options Its patented compressor technology reduces vibra tions providing comprehensive protec tion for your samples reddot design award winner 2011 And let s not forget the reliable tem perature accuracy the preset tempera ture of 4 C r
37. ntrol bands Bacillus subtilis were fully visible and the findings were confirmed by surrounding clinical re sults The analytical sensitivity of the assay was tested before to be lt 10 colony forming units cfu mL E feacium and vancomycin A resistance respectively Discussion The combination of up to 25 primer pairs in a single multiplex PCR reaction is considered to be a challenge regarding sensitivity and specificity For pathogen detection in body fluids e g sepsis causative pathogens in whole blood assays have to cover a capacious panel of targets to yield required clinical utility Page 6 While broad range primers are under ongoing discussion with respect to high numbers of false positives and the risk of contamination 10 usage of specific primers in combination seems to be an alternative if clinical sensitivity meets the range of diagnostic relevance and if maximum specificity is given The presented results of the VYOO PCR detection kit indicate that nucleic acids trace detection is possible applying high plex grades with desired sensitivity and specificity The Mastercycler pro fulfills the demanding requirements of such a complex PCR assay this thermal cycler helps to minimize the technical variance and contributes to a high process reli ability in the PCR workflow of such a sensitive and specific assay Analysis of whole blood samples by multiplex PCR for bacterial and fungal DNA is a reliable and r
38. osable laboratory plasticware Science 2008 322 5903 917 2 McDonald GR Kozuska JL Holt A Bioactive Leachates from Lab Plastics G I T Laboratory Journal Europe 2009 13 24 26 3 Lewis LK Robson M Vecherkina Y Ji C Beall G Interference with spectrophotometric analysis of nucleic acids and proteins by leaching of chemicals from plastic tubes BioTechniques 2010 48 4 297 302 More on Eppendorf Safe Lock Tubes at www eppendorf com consumables Readers service Eppendorf Safe Lock Tubes Ref no 8 Eppendorf BioPhotometer plus Ref no 221 BN 35 July 2011 Asia Pacific Tel 60 3 8023 6869 E Mail support_asiapacific eppendorf com Eppendorf BioNews Application Notes Removal of Fluorescently labeled Sensory neurosecretory Cells from Forebrain of Transgenic Medaka Embryos Using Eppendorf PatchMan NP 2 and CellTram vario and Rowiak CellSurgeon Kristin Tessmar Raible Katharina Schipany and Sabine Przemeck 1Max F Perutz Laboratories University of Vienna Austria Abstract For a representative analysis of cell clus ters in transgenic small animal models cell material with the least possible contamination by nondescript cells is desirable Here we report the successful removal of fluorescently labeled sensory neurosecretory cells from the forebrain of transgenic Medaka embryos combin ing laser assisted excision CellSurgeon Rowiak GmbH and extraction of excised cells using CellTr
39. pette holders and a pipette pen 2nd to 5th prize 1 MP3 player each 6th to 15th prize 200 bonus ep points each ACROSS DOWN 1 Opposite of cold 28 City in 26 across 1 Room or building with tools or 21 Geographic term 5 Pub tavern 29 High throughput abbr machinery 22 Genus of yeast 7 Crowd pack gang comes ina 30 ISO country code of Pakistan 2 Chemical symbol for argon 25 Visible result of centrifugation flash nowadays 31 Business contract bargain 3 Spanish word for river 27 Collection of maps 10 The first of its kind no fake 32 French salt 4 Chemical symbol for magnesium 32 Slender thin 12 Complements or completes 33 French future 5 Barium abbr 34 The direction opposite of the gamy poly and tony 35 Loud laughter on the web 6 Special computer key zenith 13 Its plural would be those 37 Marking that confirms a product s 7 Drive engine 36 Front part of the head 15 1000 metres abbr compliance with EU legislation 8 Ontario abbr 37 Poncho sleeveless garment 18 Measures and indicates the time 38 Chemical symbol for aluminum 9 Piece of literature 39 Logarithm abbr 20 If this is good you feel 40 Smallest particle of an element 11 Postal abbreviation for Illinois 41 Lyrical verse comfortable 42 Famous cape in Massachussetts 12 Southern European country in the 43 Perform carry out execute 23 Something John Volta is missing USA Mediterranean Sea 44 In the event that conj 24 High definition high density abbr 44 In
40. rity of the science classroom AAAS is working hard to keep evolution in the classroom by publishing op eds educating school boards consulting with state and local governments and by providing support for other pro evolution organisations As a AAAS member your dues support these efforts If you re not yet a AAAS member join us Together we can make a difference To learn more visit aaas org plusyou evolution ABN 3513020
41. rotor Example 4 x 1 1 kg means weights of each bucket adapter tubes filled with sample must not exceed 1 1 kg Take note of the maximum g force specified for the tubes you are using Weight class inscription on bucket 6 BioNews No 35 Safe and Efficient Centrifugation Straight from the lab Choose correct adapter Adapters must support tubes securely The tube should fit tightly into the adapter a a Swing bucket rotors e Bucket and adapter loading must be symmetric and balanced right e Position tubes in buckets in such a way that rotor pivots are stressed evenly Wrong use of adapter Correct use of adapter no secure support of upper part e Always have all 4 swing buckets on rotor even though only 2 might be in use For conical tubes use conical tube adapter with conical base Maintenance of Centrifuges Prevention is Better than Restoring This poster does not replace the operating manual For tubes with round bottom use tube adapter with flat base and rubber mat Conical tube adapter Round bottom tube adapter conical base flat base with rubber mat For your safety Automatic imbalance detection Eppendorf centrifuge families 58x 57xx 5430 R and 5417 R have imbalance sensors to prevent spinning if the samples are not properly balanced This protects device sample and user by preventing a rotor crash Check with your provider for latest updates
42. rth up to 500 For detailed guidelines about contents and format please visit If you have further questions please contact our Application Support colleagues by e mail to support eppendorf com or give them a call at 0049 1803 666789 gt BioNews No 35 News AXEL JAHNS EPPENDORF AG Success through Competence Sustained sales growth Asia gaining in significance and an operating profit that was increased by 20 1 percent all this characterizes Eppendorf s continued successful business performance in 2010 After the upheaval of the economic crisis of 2009 the economy was not yet back on solid footing in 2010 In spite of this Eppendorf can look back ona successful year based on strong growth in Asia and the recovery of the European and US economies comments Klaus Fink CEO of Eppendorf AG who swapped the Chair of the Executive Board with the Chair of the Supervisory Board on May 1 2011 On the same day Dr Dirk Ehlers succeeded him as CEO Eppendorf s success has many good reasons The excellent image in the life science industry the customers the product quality and the competence of the em ployees these four pillars of Eopendorf s continued success are examined in more detail in the 2010 Annual Report Order your personal copy now using the reply fax form on page 15 or check it out on our website New Brunswick Catalog 2011 New Brunswick Scientific joined the Eppendorf group in 2007
43. sample disruption the lytic process that is enhanced by incubation at higher temperatures can be performed online and further reduces the need for hands on time This incubation step is instru mental in delivering higher yield and purity DNA The steps of DNA binding washing drying and elution are fully supported by the epMotion TMX system delivering the final ready to use eluted high quality high yield DNA for enhanced perform The automated procedure starts with the transfer of the sample tubes to posi tion TMX followed by the addition of Lysis Buffer to each of the sample lysis tubes TMX followed by the addition of plant neutralisation buffer The lysis tubes are removed from the epMotion and are homogenised using a mixing mill Thereafter the tubes are centri fuged to remove any debris and re eae turned to the epMotion ance in downstream applications such as restriction digestion The automated protocol continues with mixing heating and incubation steps Before adding the solution of Magnetic bead amp RNase the magnetic bead sus Materials and methods Eppendorf eoMotion TMX ension should be mixed carefully The Nucleon Plant DNA Kit Gen Probe Life gt d binding and washing steps are continued Sciences San Diego CA USA 9 deep The final sample is transferred to the elution plate The total processing time is approximately 4 h for 96 samples of which 3 h are automated processes Multi Magne
44. t July 2011 BN 35 and spread of antibiotic resistances are serious health risks which are mainly contributed to antibiotics overuse 7 Compared to the so far routinely used culture based methods nucleic acid amplification techniques NAT allow a more rapid species and resistance detection within several hours Possible impact on sensitivity caused by factors like e g salts and blood ingredients can be abolished by affinity chromatography sample preparation VYOO CE marked 2008 combines such a sample prepara tion with sensitive multiplex PCR based pathogen detection and a time to result of less than one working day 8 9 Materials and methods Mechanical cell lysis EDTA whole blood was homogenized and poured into tube containing glass bead matrix antifoam solution and Bacillus subtilis endospores as internal run control Mechanical impact was applied using a FastPrep 24 cell lysis device MP Biomedicals Solon OH USA followed by proteolytic digestion of the lysate 11 Total genomic DNA isolation DNA of digested lysates was isolated by applying a standard protocol and dissolved in an appropriate buffer for subsequent affinity chromatography 11 Enrichment of bacterial and fungal DNA with LOOXSTER Total DNA was subjected onto affinity chromatography spin columns and after standard processing dissolved in 30 uL water DNA concentration was determined by photometry 11 Multiplex PCR detection
45. ternet Protocol abbr 14 Honoris causa abbr 45 Personal computer abbr 25 This US state abbr has 45 Postscript 16 Japanese rice fish 46 ISO country code of Sweden Harrisburg as its capital 47 lin Latin 17 Scanning device e g for bar 26 State on the West coast of the 48 Popular one button pipette codes or cards USA abbr Stores data Disclaimer and Trademarks Information CellTram epGreen Logo epMotion Eppendorf Eppendorf BioPhotometer Eppendorf LoBind Eppendorf Reference Eppendorf Tubes Eppendorf twin tec Eppendorf Xplorer ep points epT 1 P S Masterclear Mastercycler Multiporator PhysioCare Concept are registered trademarks of Eppendorf AG epGreen Eppendorf Safe Lock epServices epServices Logo FastTemp Kit Rotor New Brunswick Logo New Brunswick Logo with Flask Design PatchMan are trademarks of Eppendorf AG OptiBowl Silence Speed Simplicity are registered trademarks of Eppendorf North America Inc USA LOOXSTER and VYOO are registered trademarks of SIRS Lab GmbH QIAGEN is a registered trademark of QIAGEN Group Nucleon is a registered trademark of Gen Probe Life Sciences Ltd Magnabot is a registered trademark of Promega Corp Errors and omissions excepted Status July 2011 ta BioNews No 35 Service To Eppendorf AG Hamburg Germany Attn Anton Janott Fax 49 40 5 3801 8 40 Please print or type no private address Company Uni
46. tic Probe Plate Promega Magnabot 96 Promega Madison WI USA Results Sample material Yield and purity of DNA Leaves of cabbage pine rose lettuce and barley and popping corn For each individual sample 30 mg of plant mate As shown in Table 1 DNA from various plant samples can be easily purified with the Nucleon Plant DNA Kit and the automated epMotion 5075 TMX system rial was placed into the provided sample lysis tubes containing the steel ball pow sees es Jo ie g uL Cabbage 30 64 2 18 1 7 109 63 21 93 31 9 Pine 30 18 2 00 4 1 26 16 5 23 35 1 Rose 30 18 1 67 2 9 61 84 12 37 13 3 Lettuce 30 18 2 23 6 4 15 96 3 18 20 7 Corn 30 16 2 14 TA 22 88 4 58 74 1 Barley 30 8 2 06 2 9 54 01 10 0 26 3 Gee 2 05 pula 11 65 Table 1 Yield and purity of DNA Variability in sample yield is not unexpected when purifying DNA from plant material this can usually be attributed to the off line sample preparation stage Application Support July 2011 BN 35 Europe International Tel 49 1803 666 789 0 09 min aus dem dt Festnetz Mobilfunk max 0 42 min E Mail support eppendorf com Page T North America Tel 1 800 645 3050 E Mail techserv eppendorf com Asia Pacific Tel 60 3 8023 6869 E Mail support_asiapacific eppendorf com Eppendorf BioNews Application Notes _ Automated Plant DNA Purification Using the Nucleon Plant DNA Kit on the epMotion 5075 TMX The method was found to deliver initial D
47. tive operation For Eppendorf intuitive operation means fast and safe handling without time consuming training Because routine processes frequently need to be mastered quickly and easily to guarantee a high level of safety in performance Therefore the Centrifuge 5424 R is equipped with a display which is easy to read and operate The device is also available in two versions with rotary knobs for quick parameter setting or an easy to clean keypad Another plus flexible rotor selection The Centrifuge 5424 R can be used with four different rotors according to individual needs e Aerosol tight rotor for 24 x 1 5 2 0 mL tubes e Kit rotor for safe centrifugation of 18 x spin columns or 1 5 2 0 mL tubes with open tube lids The raised edge of the Kit rotor prevents open tube lids from breaking off during centrifugation e Aerosol tight rotor for 24 x 1 5 2 0 mL tubes with PTFE coating for increased chemical resistance e 4x PCR strip rotor Kit rotor F 45 18 11 Kit Our contribution to the environment epGreen Eppendorf demonstrates its commit ment to a green planet with the epGreen initiative whose objective is to constant ly reduce the environmental impact of our business operations and products Therefore the Centrifuge 5424 R was developed according to the latest emis sions and environmental requirements while also setting new standards in these areas The baseline power consumption has been re
48. ucleon Plant DNA Kit on the epMotion 5075 TMX from Eppendorf www eppendorf com applications Nucleon Plant DNA Kit user manual www gen probe com pdf Operation Manual for epMotion 5075 www eppendorf com Support amp Services Literature Library Manuals Readers service epMotion 5075 TMX e Ref no 228 BN 35 July 2011 Asia Pacific Tel 60 3 8023 6869 E Mail support_asiapacific eppendorf com Innovation CARSTEN BUHLMANN EPPENDORF AG New Box System for epT 1 P S Motion epT I P S Motion pipette tips for the epMotion automatic pipetting systems are now available in color coded trays In addition we ve optimized the packaging system With the new box system you now have a flexible solution that perfect ly adapts to your needs New and different The new ep T 1 P S Motion Rack con sists of two components a colored tray for easy tip volume identification and a box to house the tray New and ecological the refill system You can make a long term contribution to the environment with our new Re loads in sealed blister packs which can be easily used with the redesigned sturdy metal TipHolder adapter These affordable alternatives to disposable boxes offer a waste reduction of up to 40 New and flexible Whatever version you choose both systems can be immediately integrated into your epMotion The existing labware files work seamlessly with the new tips The
49. versity Clinic Title Name Institute Address Department Function Address E Mail Telephone Telefax NEW Visit www eppendorf com bn service to request literature online Please send me information on the products denoted by the following reference numbers Kindly help us to update your subscription details 8 Eppendorf Safe Lock Tubes am already an Eppendorf BioNews subscriber 30 Eppendorf Reference Please change my details See above 52 PatchMan am a new subscriber Please send me Eppendorf BioNews 77 CellTram on a regular basis 2 issues per year free of charge O OOO 180 epT P S Motion am interested in an electronic Eppendorf Newsletter Please enter your e mail address in the address field 221 Eppendorf BioPhotometer plus 223 real time PCR Consumables I have comments on the BioNews No 35 224 Mastercycler pro 228 epMotion 5075 TMX 235 Centrifuge 5424 R Eppendorf General Catalog 2011 OOOO I have suggestions topics for the next BioNews Eppendorf Q Annual Report 2010 C New Brunswick Catalog 2011 Solution of BioNews No 35 prize competition et tte Ey EL tee TE e l Closing date 31st October 2011 Visit www eppendorf com bn service to participate online BioNews No 35 GH AAAS is here protecting the integ
50. yo once cell clusters had been identified since fine movements of the microcapillary were considerably ham pered and slowed by the viscous methyl cellulose Mounting of embryos Chorionated embryos were anaesthe tised with 0 4 Tricaine and mounted onto the glass bottom of a 35 mm dish partially filled with 6 methyl cellulose Once embryos had been carefully orien tated with eyes facing the glass bottom they were screened for fluorescent cell clusters Fig 1 Embryos that had been screened positively were sacrificed with an overdose of Tricaine methansulfonate and manually hatched under a stereo microscope Hatched dechorionated embryos were also mounted using 6 methyl cellulose but placed on their backs with heads close to the dish bottom Page 3 Asia Pacific Tel 60 3 8023 6869 E Mail support_asiapacific eppendorf com f Eppendorf BioNews Application Notes Removal of Fluorescently labeled Sensory neurosecretory Cells from Forebrain of Transgenic Medaka Embryos Using Eppendorf PatchMan NP 2 and CellTram vario and Rowiak CellSurgeon Fig 1 Positioning of microcapillary Using 20x magnification and brightfield imaging the microcapillary was brought into focal plane in close proximity of the embryo s head Magnification was changed to 40x for precise positioning and rechecked using fluorescence imag ing to establish an exact position of tip in comparison to cell cluster This position was stored

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