NucleoMag® Trace - MACHEREY
Contents
1. Genomic DNA from forensic samples User manual NucleoMag Trace August 2015 Rev 05 MACHEREY NAGEL MN www mn net com Genomic DNA from forensic samples Table of contents 1 Components 4 1 1 Kit contents 4 1 2 Consumables and equipment to be supplied by user 5 2 Product description 6 2 1 The basic principle 6 2 2 Kit specifications 6 2 3 Magnetic separation systems 7 2 4 Adjusting the shaker settings 8 2 5 Handling of beads 8 2 6 Elution procedures 9 3 Storage conditions and preparation of working solutions 10 4 Safety instructions 11 5 Protocol for the isolation of genomic DNA from forensic samples 13 6 Appendix 18 6 1 Troubleshooting 18 6 2 Ordering information 20 6 3 Product use restriction warranty 21 MACHEREY NAGEL 08 2015 Rev 05 3 Genomic DNA from forensic samples 1 Components 1 1 Kit contents NucleoMag Trace 1x 96 preps 4 x 96 preps 24 x 96 preps REF 744600 1 744600 4 744600 24 NucleoMag B Beads 1 7 mL 7 mL 42 mL Lysis Buffer FLB 50 mL 250 mL 2 x 500 mL Binding Buffer MB2 45 mL 180 mL 2 x 480 mL Wash Buffer MB3 75 mL 300 mL 2 x 900 mL Wash Buffer MB4 75 mL 300 mL 2 x 900 mL Wash Buffer MB5 125 mL 500 mL 3 x 1000 mL Elution Buffer MB6 30 mL 125 mL 2 x 300 mL Proteinase K lyophilized 50 mg 4x 50 mg 24 x 50 mg Proteinase Buffer PB 8 mL 15 mL 3x 35 mL User manual 1 1 1 For preparation of working solutions and storage conditions see section 3 4 MACHEREY NAGE2. Genomic DNA from forensic samples 6 Appendix 6 1 Troubleshooting Problem Possible cause and suggestions Elution buffer volume insufficient Beads pellet must be covered completely with elution buffer Insufficient performance of elution buffer during elution step Remove residual buffers during the separation steps completely Remaining buffers decrease efficiency of following wash steps and elution step Beads dried out Do not let the beads dry as this might result in lower elution efficiencies Partial elution in Wash Buffer MB5 already Keep the beads on the magnet while dispensing Wash Poor DNA Buffer MB5 Do not resuspend beads in this buffer and do yield not incubate beads in this buffer for more than 2 min as this buffer is water based and might elute the DNA already Aspiration of attracted bead pellet Do not disturb the attracted beads while aspirating the supernatant especially when the magnetic pellet is not visible in the lysate Incubation after dispensing beads to lysate Mix immediately after dispensing NucleoMag B Beads Buffer MB2 to the lysate Aspiration and loss of beads Time for magnetic separation was too short or aspiration speed was too high Insufficient washing procedure Use only the appropriate combinations of separator and plate for example Square well Block in combination with NucleoMag SEP Low purity Make sure that beads are resuspended completely during the washing procedure If sha
3. MACHEREY NAGEL 08 2015 Rev 05 13 NucleoMag Trace After 2 min separation remove supernatant 5 Wash with MB4 Remove Square well Block from NucleoMag SEP 600 uL MB4 Resuspend Shake 5 min at RT Optional Mix by pipetting up and down After 2 min separation remove supernatant 6 Wash with MB5 Leave Square well Block on NucleoMag SEP 900 uL MB5 Incubate for 45 60 s Note Do not resuspend the beads in Buffer MB5 Remove supernatant 7 Elute DNA Remove Square well Block from NucleoMag SEP 50 200 pL MB6 Optional Elute at 56 C Shake 5 min at RT Optional Mix by pipetting up and down Separate 2 min and transfer DNA into elution plate tubes 14 MACHEREY NAGEL 08 2015 Rev 05 NucleoMag Trace Detailed protocol This protocol is designed for magnetic separators with static pins e g NucleoMag SEP and suitable plate shakers see section 2 3 It is recommended using a Square well Block for separation see section 1 2 Alternatively isolation of DNA can be performed in reaction tubes with suitable magnetic separators This protocol is for manual use and serves as a guideline for adapting the kit to robotic instruments Before starting the preparation Check if Proteinase K was prepared according to section 3 Sample collection Collect the samples with cotton Dacron or C E P swabs Scrape firmly against
4. the inside of each cheek several times and let the swabs air dry The respective individual should not have consumed food or drink within 30 min before collection of the sample Samples should be processed immediately or stored at 4 C 1 Lyse samples Calculate the amount of lysis stock required for each sample 25 uL of Proteinase K solution 200 uL Buffer FLB are required Prepare lysis stock solution accordingly and vortex Note Never prepare the lysis stock solution more than 15 min before addition to the samples Proteinase K tends to self digestion when incubated in Buffer FLB without substrate Transfer 225 uL of the resulting solution to each lysis tube containing the buccal swab head Close the individual tubes Mix by vigorous shaking for 10 15 s Spin briefly 15 s 1 500 x g to collect any sample at the bottom of the tube Note The buccal swab heads should be submerged into the lysis solution Therefore depending on type or size of buccal swab used the FLB buffer volume has to be increased to up to 400 uL Increasing volume of Proteinase K solution is not required Alternatively perform lysis with Buffer FLB Proteinase K in a NucleoSpin Trace Filter Plate see ordering information This plate allows convenient separation of lysate from swab material by centrifugation and reduces loss of lysate Incubate the tubes containing the samples at 56 C for 1 h or overnight at room temperature For optimal lysis m
5. bottle Vortex storage bottle briefly until a homogenous suspension has formed Separate the magnetic beads against the side of the wells by placing the Square well Block on the NucleoMag SEP magnetic separator Wait at least 2 min until all the beads have been attracted to the magnets Remove and discard supernatant by pipetting Note Do not disturb the attracted beads while aspirating the supernatant The magnetic pellet is not visible in this step Remove supernatant from the opposite side of the well Wash with MB3 Remove the Square well Block from the NucleoMag SEP magnetic separator Add 600 uL Buffer MB3 to each well and resuspend the beads by shaking until the beads are resuspended completely 5 min Alternatively resuspend beads completely by repeated pipetting up and down 15 times 16 MACHEREY NAGEL 08 2015 Rev 05 NucleoMag Trace Separate the magnetic beads by placing the Square well Block on the NucleoMag SEP magnetic separator Wait at least 2 min until all the beads have been attracted to the magnet Remove and discard supernatant by pipetting Wash with MB4 Remove the Square well Block from the NucleoMag SEP magnetic separator Add 600 uL Buffer MB4 to each well and resuspend the beads by shaking until the beads are resuspended completely 5 min Alternatively resuspend beads completely by repeated pipetting up and down 15 times Separate the magnetic beads by placing the Squar
6. the plate surface for small droplets of dyed water Increase speed setting shake for an additional 30 seconds and check the plate surface for droplets again Continue increasing the speed setting until you observe droplets on top of the separation plate Reduce speed setting check again and use this setting for the washing step Adjusting shaker speed for the elution step Load 100 200 uL dyed water to the wells of the collection plate and proceed as described above 2 5 Handling of beads Distribution of beads A homogeneous distribution of the magnetic beads to the individual wells of the separation plate is essential for a high well to well consistency Therefore before distributing the beads make sure that the beads are completely resuspended Shake the storage bottle well or place it on a vortexer shortly Premixing magnetic beads with the binding buffer allows easier homogenous distribution of the beads to the individual wells of the separation plate During automation a premix step before aspirating the beads binding buffer mixture from the reservoir is recommended to keep the beads resuspended Magnetic separation time Attraction of the magnetic beads to the magnetic pins depends on the magnetic strength of the magnetic pins the selected separation plate distance of the separation plate from the magnetic pins and the volume to be processed The individual times for complete attraction of the beads to the magneti
7. EL literature are provided for informational purposes only MACHEREY NAGEL does not warrant that all applications have been tested in MACHEREY NAGEL laboratories using MACHEREY NAGEL products MACHEREY NAGEL does not warrant the correctness of any of those applications Last updated 07 2010 Rev 03 Please contact MACHEREY NAGEL GmbH amp Co KG Tel 49 24 21 969 270 tech bio mn net com Trademarks KingFisher is a registered trademark of Thermo Fisher Scientific NucleoMag is a registered trademark of MACHEREY NAGEL GmbH amp Co KG Te MagS is a trademark of Tecan Group Ltd Switzerland All used names and denotations can be brands trademarks or registered labels of their respective owner also if they are not special denotation To mention products and brands is only a kind of information i e it does not offend against trademarks and brands and can not be seen as a kind of recommendation or assessment Regarding these products or services we can not grant any guarantees regarding selection efficiency or operation 22 MACHEREY NAGEL 08 2015 Rev 05 MACHEREY NAGEL MACHEREY NAGEL GmbH amp Co KG Neumann Neander Str 6 8 52355 D ren Germany Germany Switzerland France USA and international MACHEREY NAGEL AG MACHEREY NAGEL EURL MACHEREY NAGEL Inc Tel 49 24 21 969 0 Tel 41 62 388 55 00 Tel 33 388 68 22 68 Tel 1 484 821 0984 E mail info mn net com E mail sales ch mn net com E m
8. L 08 2015 Rev 05 Genomic DNA from forensic samples 1 2 Consumables and equipment to be supplied by user Product REF Magnetic separator 744900 NucleoMag SEP see section 2 3 Separation plate for magnetic beads separation 740481 Square well Block 96 well block with 2 1 mL square 740481 24 wells Lysis tubes for incubation of samples and Iysis 740477 e g Rack of Tubes Strips 1 set consists of 1 Rack 740477 24 12 Strips with 8 tubes 1 2 mL wells each and 12 Cap Strips Elution plate for collecting purified nucleic acids 740486 24 e g Elution Plate U bottom 96 well 0 3 mL microtiterplate with 300 uL u bottom wells For use of kit on KingFisher 96 instrument 744950 e g KingFisher 96 Accessory Kit A Square well Blocks Deep well tip combs Plates for 4 x 96 NucleoMag Trace preps using KingFisher 96 platform Pack of 1 4 24 4 sets 24 sets 24 1 set MACHEREY NAGEL 08 2015 Rev 05 Genomic DNA from forensic samples 2 Product description 2 1 The basic principle The NucleoMag Trace procedure is based on reversible adsorption of nucleic acids to paramagnetic beads under appropriate buffer conditions Lysis is achieved by incubation of samples with Proteinase K at room temperature or 56 C For the adjustment of binding conditions under which nucleic acids bind to the paramagnetic beads Buffer MB2 and the NucleoMag B Beads are added to the lysate After ma
9. P261 P272 P280 P301 312 P302 352 P304 340 P330 P333 313 P342 311 P370 378 P363 P403 235 Avoid breathing dust fume gas mist vapours spray Einatmen von Staub Rauch Gas Nebel Dampf Aerosol vermeiden Contaminated work clothing should not be allowed out of the workplace Kontaminierte Arbeitskleidung nicht au erhalb des Arbeitsplatzes tragen Wear protective gloves protective clothing eye protection face protection Schutzhandschuhe Schutzkleidung Augenschutz Gesichtsschutz tragen IF SWALLOWED Call a POISON CENTER doctor if you feel unwell BEI VERSCHLUCKEN Bei Unwohlsein GIFTINFORMATIONSZENTRUM Arzt anrufen IF ON SKIN Wash with plenty of water BEI BERUHRUNG MIT DER HAUT Mit viel Wasser waschen IF INHALED Remove person to fresh air and keep comfortable for breathing BEI EINATMEN Die Person an die frische Luft bringen und f r ungehinderte Atmung sorgen Rinse mouth Mund aussp len If skin irritation or rash occurs Get medical advice attention Bei Hautreizung oder ausschlag Arztlichen Rat einholen rztliche Hilfe hinzuziehen If experiencing respiratory symptoms Call a POISON CENTER doctor Bei Symptomen der Atemwege GIFTINFORMATIONSZENTRUM Arzv anrufen In case of fire Use to extinguish Bei Brand zum L schen verwenden Wash contaminated clothing before reuse Kontaminierte Kleidung vor erneutem Tragen waschen Store in a well ventilated place Ke
10. Y CONDITION OR ANY OTHER MATTER WITH RESPECT TO MACHEREY NAGEL PRODUCTS In no event shall MACHEREY NAGEL be liable for claims for any other damages whether direct indirect incidental compensatory foreseeable consequential or special including but not limited to loss of use revenue or profit whether based upon warranty contract tort including negligence or strict liability arising in connection with the sale or the failure of MACHEREY NAGEL products to perform in accordance with the stated specifications This warranty is exclusive and MACHEREY NAGEL makes no other warranty expressed or implied The warranty provided herein and the data specifications and descriptions of this MACHEREY NAGEL product appearing in MACHEREY NAGEL published catalogues and product literature are MACHEREY NAGEL s sole representations concerning the product and warranty No other statements or representations written or oral by MACHEREY NAGEL s employees agent or representatives except written statements signed by a duly authorized officer of MACHEREY NAGEL are authorized they should not be relied upon by the customer and are not a part of the contract of sale or of this warranty Product claims are subject to change Therefore please contact our Technical Service Team for the most up to date information on MACHEREY NAGEL products You may also contact your local distributor for general scientific information Applications mentioned in MACHEREY NAG
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12. c pins should be checked and adjusted on each system It is recommended using the separation plates or tubes specified by the supplier of the magnetic separator 8 MACHEREY NAGEL 08 2015 Rev 05 Genomic DNA from forensic samples Washing the beads Washing the beads can be achieved by shaking or mixing In contrast to mixing by pipetting up and down mixing by shaker or magnetic mixing allows simultaneous mixing of all samples This reduces the time and number of tips needed for the preparation Resuspension by pipetting up and down however is more efficient than mixing by a shaker or magnetic mix Resuspension Small elution Number of tips is Speed f efficiency volume possible needed Magnetic mix Low Shaker Low Pipetting High acceptable good excellent 2 6 Elution procedures Purified DNA can be eluted directly with the supplied Elution Buffer MB6 Elution can be carried out in a volume of gt 50 uL It is essential to cover the NucleoMag Beads completely with elution buffer during the elution step The volume of dispensed elution buffer depends on the magnetic separation system e g the position of the pellet inside the separation plate For efficient elution the magnetic bead pellet should be resuspended completely in the elution buffer For some separators higher elution volumes might be necessary to cover the whole pellet Elution is possible at room t
13. e well Block on the NucleoMag SEP magnetic separator Wait at least 2 min until all the beads have been attracted to the magnet Remove and discard supernatant by pipetting Wash with MB5 Leave the Square well Block on the NucleoMag SEP magnetic separator Note Supernatant is colorless magnetic bead pellet is clearly visible Gently add 900 uL Buffer MB5 to each well and incubate for 45 60 s while the beads are still attracted to magnets Then aspirate and discard the supernatant Note Do not resuspend the beads in Wash Buffer MB5 This step is to remove traces of ethanol and eliminates a drying step Elute DNA Remove the Square well Block from the NucleoMag SEP magnetic separator Add desired volume of Buffer MB6 50 200 uL to each well of the Square well Block and resuspend the beads by shaking 5 10 min at room temperature or 56 C Alternatively resuspend beads completely by repeated pipetting up and down and incubate for 5 10 min at room temperature or 56 C Separate the magnetic beads by placing the Square well Block on the NucleoMag SEP magnetic separator Wait at least 2 min until all the beads have been attracted to the magnets Transfer the supernatant containing the purified genomic DNA to the Elution Plate Note Yield can be increased by 15 20 by using pre heated elution buffer 56 C or by incubating the bead elution buffer suspension at 56 C for 10 min MACHEREY NAGEL 08 2015 Rev 05 17
14. emperature Yield can be increased by 15 20 if elution is performed at 56 C MACHEREY NAGEL 08 2015 Rev 05 9 Genomic DNA from forensic samples 3 Storage conditions and preparation of working solutions Attention Buffers MB2 MB3 and MB4 contain chaotropic salt Wear gloves and goggles Storage conditions All components of the NucleoMag Trace kit should be stored at room temperature 18 25 C and are stable for at least one year All buffers are delivered ready to use Before starting any NucleoMag Trace protocol prepare the following Proteinase K Add the indicated volume of Proteinase Buffer PB to dissolve lyophilized Proteinase K Proteinase K solution is stable at 20 C for at least 6 months NucleoMag Trace 1 x 96 preps 4x 96 preps 24 x 96 preps REF 744600 1 744600 4 744600 24 Proteinase K 1x 50 mg 4x50 mg 24 x 50 mg lyophilized Add 2 5 mL Add 2 5 mL Add 2 5 mL Proteinase Buffer Proteinase Buffer to Proteinase Buffer each vial to each vial 10 MACHEREY NAGEL 08 2015 Rev 05 Genomic DNA from forensic samples 4 Safety instructions The following components of the NucleoMag Trace kits contain hazardous contents Wear gloves and goggles and follow the safety instructions given in this section GHS classification Only harmful features do not need to be labeled with H and P phrases up to 125 mL or 125 g Mindergef hrliche Eigenschaften m ssen bis 125 mL oder 125 g nicht
15. ep cool An einem gut bel fteten Ort aufbewahren K hl halten For further information please see Material Safety Data Sheets www mn net com Weiterf hrende Informationen finden Sie in den Sicherheitsdatenbl ttern www mn net com A The symbol shown on labels refers to further safety information in this section Das auf Etiketten dargestellte Symbol weist auf weitere Sicherheitsinformationen dieses Kapitels hin 12 MACHEREY NAGEL 08 2015 Rev 05 NucleoMag Trace 5 Protocol for the isolation of genomic DNA from forensic samples Protocol at a glance For additional equipment and hardware requirements refer to section 1 2 and 2 3 respectively For detailed information on each step see page 15 Before starting the preparation Check if Proteinase K was prepared according to section 3 1 Lyse sample Add 25 uL Proteinase K solution e g buccal swabs and 200 400 uL Buffer FLB Mix 56 C 1h 2 Separate lysate from sample material transfer 225 uL of lysate toa Square well Block for further processing 3 Bind DNAto 225 uL lysate NucleoMag B Beads 14 pL NucleoMag B Beads 360 pL MB2 Mix by shaking for 5 min at RT e gt Optional Mix by pipetting up and down After 2 min separation remove supernatant 4 Wash with MB3 Remove Square well Block from NucleoMag SEP 600 pL MB3 Resuspend Shake 5 min at RT Optional Mix by pipetting up and down
16. f the product MACHEREY NAGEL products shall exclusively be used in an ADEQUATE TEST ENVIRONMENT MACHEREY NAGEL does not assume any responsibility for damages due to improper application of our products in other fields of application Application on the human body is STRICTLY FORBIDDEN The respective user is liable for any and all damages resulting from such application DNA RNA PROTEIN purification products of MACHEREY NAGEL are suitable for IN VITRO USES ONLY ONLY MACHEREY NAGEL products specially labeled as IVD are also suitable for IN VITRO diagnostic use Please pay attention to the package of the product IN VITRO diagnostic products are expressly marked as IVD on the packaging IF THERE IS NO IVD SIGN THE PRODUCT SHALL NOT BE SUITABLE FOR IN VITRO DIAGNOSTIC USE ALL OTHER PRODUCTS NOT LABELED AS IVD ARE NOT SUITED FOR ANY CLINICAL USE INCLUDING BUT NOT LIMITED TO DIAGNOSTIC THERAPEUTIC AND OR PROGNOSTIC USE No claim or representations is intended for its use to identify any specific organism or for clinical use included but not limited to diagnostic prognostic therapeutic or blood banking It is rather in the responsibility of the user or in any case of resale of the products in the responsibility of the reseller to inspect and assure the use of the DNA RNA protein purification products of MACHEREY NAGEL for a well defined and specific application MACHEREY NAGEL shall only be responsible for the product specificat
17. gnetic separation the paramagnetic beads are washed twice to remove contaminants and salts using Wash Buffers MB3 and MB4 Residual ethanol from previous wash steps is removed by Wash Buffer MB5 Finally highly purified DNA is eluted with low salt Elution Buffer MB6 and can directly be used for downstream applications The NucleoMag Trace kit can be used either manually or automated on standard liquid handling instruments or automated magnetic separators 2 2 Kit specifications NucleoMag Trace is designed for rapid manual and automated small scale preparation of highly pure genomic DNA from buccal swabs or other samples for example dried blood spots or cigarette filters The kit is designed for use with NucleoMag SEP magnetic separator plate see ordering information or other magnetic separation systems see section 2 3 Manual time for the preparation of 96 samples is about 120 minutes The purified DNA can be used directly as template for PCR or any kind of enzymatic reactions NucleoMag Trace allows easy automation on common liquid handling instruments or automated magnetic separators The actual processing time depends on the configuration of the instrument and the magnetic separation system used Typically 96 samples can be purified in less than 120 minutes using the NucleoMag SEP on the automation platform The kit provides reagents for the purification of up to 7 ug of pure genomic DNA from suitable samples typica
18. in the buffer by pipetting up and down several times For fully automated use on liquid handling workstations a gripper tool is required the plate is transferred to the magnetic separator for separation of the beads and transferred to the shaker module for resuspension of the beads Movable magnetic systems Separators with moving magnetic pins Magnetic pins rods are moved from one side of the well to the other and vice versa Beads follow this movement and are thus pulled through the buffer during the wash and elution steps Separation takes place when the system stops Automated separators Separators with moving magnets Magnetic beads are transferred into suitable plates or tubes Beads are resuspended from the rod covered magnets Following binding washing or elution beads are collected again with the rod covered magnets and transferred to the next plate or tube MACHEREY NAGEL 08 2015 Rev 05 7 Genomic DNA from forensic samples 2 4 Adjusting the shaker settings When using a plate shaker for the washing and elution steps the speed settings have to be adjusted carefully for each specific separation plate and shaker to prevent cross contamination from well to well Proceed as follows Adjusting shaker speed for binding and wash steps Load 600 uL dyed water to the wells of the separation plate Place the plate on the shaker and start shaking with a moderate speed setting for 30 seconds Turn off the shaker and check
19. ions and the performance range of MN products according to the specifications of in house quality control product documentation and marketing material This MACHEREY NAGEL product is shipped with documentation stating specifications and other technical information MACHEREY NAGEL warrants to meet the stated specifications MACHEREY NAGEL s sole obligation and the customer s sole remedy is limited to replacement of products free of charge in the event products fail to perform as warranted Supplementary reference is made to the general business terms and conditions of MACHEREY NAGEL which are printed on the price list Please contact us if you wish to get an extra copy There is no warranty for and MACHEREY NAGEL is not liable for damages or defects arising in shipping and handling transport insurance for customers excluded or out of accident or improper or abnormal use of this product defects in products or MACHEREY NAGEL 08 2015 Rev 05 21 Genomic DNA from forensic samples components not manufactured by MACHEREY NAGEL or damages resulting from such non MACHEREY NAGEL components or products MACHEREY NAGEL makes no other warranty of any kind whatsoever and SPECIFICALLY DISCLAIMS AND EXCLUDES ALL OTHER WARRANTIES OF ANY KIND OR NATURE WHATSOEVER DIRECTLY OR INDIRECTLY EXPRESS OR IMPLIED INCLUDING WITHOUT LIMITATION AS TO THE SUITABILITY REPRODUCTIVITY DURABILITY FITNESS FOR A PARTICULAR PURPOSE OR USE MERCHANTABILIT
20. ix occasionally during incubation Make sure that the lysis tubes are securely closed Note Other samples e g dried blood spots cigarette filters can be processed accordingly MACHEREY NAGEL 08 2015 Rev 05 15 NucleoMag Trace Separate lysate Separate swab material from lysed sample Remove buccal swab and squeeze out to obtain 225 uL lysate When using increased volumes gt 200 uL of Buffer FLB in step 1 of the procedure transfer 225 uL lysed sample to a new Square well Block for further processing When using the NucleoSpin Trace Filter Plate centrifuge the NucleoSpin Trace Filter Plate stacked onto a 96 well Square well Block for 5 min at 5 600 x g to draw the lysate out of the swab material Bind DNA to NucleoMag B Beads To each lysate of 225 uL from the previous step add 14 pL of NucleoMag B Beads and 360 uL of Binding Buffer MB2 Mix by pipetting up and down 6 times and shake for 5 min at room temperature Alternatively when processing the kit without a shaker pipette up and down 10 times and incubate for 5 min at room temperature Note NucleoMag B Beads and Buffer MB2 can be premixed Per well to be processed mix 14 uL of NucleoMag B Beads with 360 uL Buffer MB2 Vortex briefly Depending on the dead volume of the reservoir additional amounts of bead suspension and binding buffer are required Be sure to resuspend the NucleoMag B Beads before removing them from the storage
21. king is not sufficient to resuspend the beads completely mix by repeated pipetting up and down 18 MACHEREY NAGEL 08 2015 Rev 05 Genomic DNA from forensic samples Problem Suboptimal performance of DNA in downstream applications Possible cause and suggestions Carry over of ethanol from wash buffers Be sure to remove all of the ethanolic wash solution as residual ethanol interferes with downstream applications Low purity See above Carry over of beads Time for magnetic separation too short Increase separation time to allow the beads to be completely attracted to the magnetic pins before aspirating any liquid from the well Aspiration speed too high elution step High aspiration speed during the elution step may cause bead carry over Reduce aspiration speed for elution step Cross contamination Contamination of the rims Do not moisten the rims of the Square well Block when transferring the lysate If the rim of the wells is contaminated seal the Square well Block with Self adhering PE Foil see ordering information before starting the shaker MACHEREY NAGEL 08 2015 Rev 05 19 Genomic DNA from forensic samples 6 2 Ordering information Product NucleoMag Trace NucleoMag Trace NucleoSpin Forensic Filters Bulk NucleoSpin Trace Filter Plate NucleoMag SEP Square well Blocks Square well Blocks ethylene oxide treated Self adhering PE F
22. l yields for DNA isolation from buccal swabs 1 3 ug DNA Depending on the elution volume used concentrations of 10 30 ng uL can be obtained Following lysis of samples with Proteinase K at 56 C recommended optional Proteinase K treatment can be performed at RT NucleoMag Trace can be processed completely at room temperature however elution at 56 C will increase the yield by about 15 20 NucleoMag B Beads are highly reactive superparamagnetic beads The binding capacity is 0 4 ug of gDNA per 1 uL of NucleoMag B Bead suspension 1 uL of suspension contains 130 ug of beads 6 MACHEREY NAGEL 08 2015 Rev 05 Genomic DNA from forensic samples 2 3 Magnetic separation systems For use of NucleoMag Trace the use of the magnetic separator NucleoMag SEP is recommended Separation is carried out in a Square well Block see ordering information The kit can also be used with other common separators UET AE LEOLE Separation plate or tube NucleoMag SEP MN REF 744900 Square well Block MN REF 740481 Tecan Te MagS 1 5 mL tubes without lid Sarstedt Static magnetic pins Separators with static magnetic pins for example NucleoMag SEP for manual use and for use on liquid handling workstations This type of separator is recommended in combination with a suitable microplate shaker for optimal resuspension of the beads during the washing and elution steps Alternatively beads can be resuspended
23. mit H und P S tzen gekennzeichnet werden GHS Hazard Precaution Component Hazard contents symbol phrases phrases Inhalt Gefahrstoff GHS Symbol H S tze P S tze MB2 Ethanol 35 55 sodium 226 302 210 233 perchlorate 20 40 301 312 330 Ethanol 35 55 370 378 Natriumperchlorat 20 40 WARNING 403 235 ACHTUNG CAS 64 17 5 7601 89 0 MB3 MB4 Ethanol 20 35 226 210 233 Ethanol 20 35 370 378 403 235 CAS 64 17 5 WARNING ACHTUNG Proteinase K Proteinase K lyophilized 317 334 261 272 280 90 100 lt gt 302 352 Proteinase K lyophilisiert 304 340 90 100 WARNING 333 313 ACHTUNG CAS 39450 01 6 9424311 969 Hazard phrases H226 Flammable liquid and vapour Fl ssigkeit und Dampf entz ndbar H302 Harmful if swallowed Gesundheitssch dlich bei Verschlucken H317 May cause an allergic skin reaction Kann allergische Hautreaktionen verursachen H334 May cause allergy or asthma symptoms or breathing difficulties if inhaled Kann bei Einatmen Allergie asthmaartige Symptome oder Atembeschwerden verursachen Precaution phrases P210 Keep away from heat hot surfaces sparks open flames and other ignition sources No smoking Von Hitze heissen Oberfl chen Funken offenen Flammen sowie anderen Z ndquellenarten fernhalten Nicht rauchen P233 Keep container tightly closed Beh lter dicht verschlossen halten MACHEREY NAGEL 08 2015 Rev 05 11 Genomic DNA from forensic samples
24. oil Rack of Tube Strips set consists of 1 Rack 12 Tube Strips with 8 tubes each and 12 Cap Strips Cap Strips KingFisher 96 Accessory Kit A set consists of Square well Blocks REF 744600 1 744600 4 744600 24 740988 10 740988 50 740988 250 740988 50B 740988 250B 740988 1000B 740677 744900 740481 740481 24 740481EO 740676 740477 740477 24 740638 744950 Deep well tip combs Elution Plates for 4 x 96 NucleoMag Trace preps using KingFisher 96 platform Visit www mn net com for more detailed product information Pack of 1 x 96 preps 4 x 96 preps 250 x 96 preps 10 x 96 pieces 50 x 96 pieces 250 x 96 pieces 50 x 96 pieces 250 x 96 pieces 1000 x 96 pieces 20 1 4 24 4 50 sheets 4 sets 24 sets 30 strips 1 set 20 MACHEREY NAGEL 08 2015 Rev 05 Genomic DNA from forensic samples 6 3 Product use restriction warranty NucleoMag Trace kit components are intended developed designed and sold FOR RESEARCH PURPOSES ONLY except however any other function of the product being expressly described in original MACHEREY NAGEL product leaflets MACHEREY NAGEL products are intended for GENERAL LABORATORY USE ONLY MACHEREY NAGEL products are suited for QUALIFIED PERSONNEL ONLY MACHEREY NAGEL products shall in any event only be used wearing adequate PROTECTIVE CLOTHING For detailed information please refer to the respective Material Safety Data Sheet o
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