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1. 6 In 1957 the first DNA polymerase was identified by Arthur Kornberg in which organism ERT7 phage J Escherichia coli IA Thermus aquaticus 7 In 1970 D Baltimore challenged the central dogma of molecular biology with which of the following EJ Taq polymerase J Reverse transcriptase Isi Restriction enzymes 0192623 80 0 9 G02 7 qQ E Q7 Q SJOMSUy 8 The following enzyme is a reverse transcriptase Eg integrase IJ Ligase 3 Telomerase 9 For one step RT PCR which primer option should be used EJ Oligo dT J Random primers Isi Sequence specific primers FJ ALl of the above 10 Why does raising the temperature in the reaction help with first strand cDNA synthesis when using thermostable reverse transcriptases EJ Raising the reaction temperature to 50 C helps alleviate complex secondary structures EJ Raising the reaction temperature speeds up the process 5x FJ Raising the reaction temperature does not help with cDNA synthesis How did you do 1 3 correct Rookie You may need a refresher course Find amazing PCR resources at lifetechnologies com elevatepcr 4 7 correct Pilot You re on your way to becoming a PCR ace Get there faster with our Real Time PCR Troubleshooting Tool at lifetechnologies com pcrsupport 8 10 correct Ace You really know your PCR Check out products to make your research even more successful at lifetechnologies com pcr disease spotlight
2. LIFE OF CELLS wv You know every cell type has its own personality so do we That s why we ve brought several cell types to life by means of clay animation in the Gibco Life of Cells video series You ll recognize these characters from your own studies Mr Jurkat Vero CHO S Stem Cell and Ms HUVEC each display their unique personality as they star in their own video WHICH CELL TYPE IS NEXT Is there a cell type that you think would look great in a clay animation video Let us know at lifetechnologies com lifeofcells Ready for a giggle Check out the Gibco Life of Cells videos at lifetechnologies com lifeofcells CHOS Stem Cell Vero Ms HUVEC So you want protein The name s Cell Stem Fey Vero Wate ou Hello my name is protein and more Cell and lm full of guys treat me poorly Ms HUVEC Some protein No problem am potential Culture LESE re Prep Wie elites people think I can be CHO S and am a protein me characterize of chemicals toxins pretty demanding powerhouse Turn to me me differentiate me parasites or even oh well you give me when you want to clone Whatever the role I m infecting me with me a what want and I ll give transfect and express your guy virus Lay it on me I can you relevant data Just take It and then some sayin NOW BECOME CERTIFIED BY THE LEADING AUTHORITY IN CELL CULTURE Learn the basic techniques for effective reproducible laboratory cell culture inc
3. life in the lab Click iT Plus TUNEL Assay for Multiplexed In Situ Detection Detects apoptotic cells in tissue and cultured cell samples through the use of a small jie highly specific labeling moiety and a bright fluorescent dye WesternDot secondary antibody conjugates Differently labeled WesternDot antibodies can be applied to a single blot for simultaneous detection of multiple proteins using standard gel or blot imaging platforms No stripping and re probing or multiple blots are required for multiple protein detection as commonly needed with ECL or colorimetric detection A Flow cytometry tutorials A Molecular Probes training videos teaching the principles of flow cytometry learn about how a flow cytometer works what kind of data can be collected and displayed by flow cytometry and much more lifetechnologies com flowtutorials BEAUTIFUL IMAGES AGAIN AND AGAIN Molecular Probes School of Fluorescence earn the fundamentals of fluorescence imaging The Molecular Probes School of Fluorescence is a comprehensive online guide where our bench scientists share the best tips tricks and protocols that will help you capture beautiful images again and again lifetechnologies com imagingbasics WORRY FREE FLUORESCENCE IMAGING Problem Weak or uneven staining especially with antibodies Possible cause Improper cell permeabilization solution The Image iT Fixation Permeabilization
4. low concentrations of cholesterol using Qubit 3 0 Fluorometer e Detects as little as 200 nM 80 ng ml cholesterol e Helps accurately measure cholesterol content of 0 01 ul of human serum e Detects both free cholesterol and cholesteryl esters e Allows for multiple time point measurements e Designed for minimal autofluorescence interference Do you run NGS Qubit 3 0 Fluorometer and Qubit dsDNA HS Assay Kit should be used to quantify the double stranded final library Qubit 3 0 NGS Starter Kit for next generation sequencing includes e One Qubit 3 0 Fluorometer e Qubit assay tubes set of 500 e Qubit dsDNA HS Assay Kit 500 assay Want to learn more Go to lifetechnologies com qubit ufetechnologies com FIND FHE AIL THAT S RID VA PURIFICATION PRODUCT It s not always easy to decide exactly which of the many available products is best suited for your specific experiment Our convenient interactive RNA product selection guide will help you find the purification product that is recommended for your sample type starting sample amount chosen purification technology and downstream application Check out and bookmark the online selection guide at lifetechnologies com rnaselection REAGENT REWARD PRESERVE AND PROTECT OLD FAITHFUL Ambion RNAlater stabilization solution Ambion TRIzol reagents You have the power to make time stand still Simply add Still the preferred solution for total RNA
5. pair you need for resequencing the human exome The Primer Designer Tool searches a database of 300 000 predesigned primer pairs to find your match and features a streamlined ordering process that allows easy online purchases See how easy finding primers pairs can be at lifetechnologies com primerdesigner Continued from page 6 This approach limits quantification to library constructs that contain both adapter sequences Ultimately using digital PCR to quantify NGS libraries decreases overall sequencing costs by ensuring an accurate quantification upfront minimizing the need to re run or repeat sequencing of samples Probe FWD Adapter Sequence of Interest E Adapter Figure 1 Schematic of TaqMan assay design for NGS library quantification on the QuantStudio 3D Digital PCR system The assay contains a forward primer reverse primer and TagMan probe all complementary to the lon Torrent or Illumina library adapter sequences Find out more at lifetechnologies com librariesquantification ufetechnologies com With Applied Biosystems and Invitrogen instruments and reagents 30 YEARS OF qPCR EXPERIENCE IN ONE BUNDLE That s why so many researchers trust SuperScript and SYBR reagents Cited and trusted by researchers SuperScript and SYBR reagents offer high specificity and sensitivity for your qRT PCR experiments Both products have been successfully used together for many applications includin
6. Cellular Dynamics International life in the lab CULTURE AND TRANSFECT ACROSS MANY CELL TYPES WITH INVITROGEN anp GIBCO REAGENTS Lipofectamine MessengerMAX reagent can transfect mesenchymal stem cells at high efficiency without the toxic effects of viral vectors uonposJsueJ Daniel Cohen PhD University of Pennsylvania Get a gt 20 fold increase in your membrane protein yield For researchers studying G protein coupled receptors GPCRs and other membrane proteins the Expi293 MembranePro Expression System offers enriched functional membrane proteins e Efficient greater than 20 fold increase in membrane protein yield compared to the adherent culture system e Scalable easy scale up without the need for multiple flasks e Easy to use no enzymatic or mechanical dissociation required lifetechnologies com membranepro ufetechnologies com Purchase Lipofectamine MessengerMAX reagent and receive a cool T shirt Lipofectamine MessengerMAX Transfection Reagent is your ticket to higher mRNA transfection efficiency Lipofectamine MessengerMAX mRNA Transfection Reagent delivers amazing transfection efficiency in neurons and a broad spectrum of primary cells enablng improved application outcomes and more biologically relevant research e Amazing transfection efficiency in neurons and primary cell types e Faster protein expression with no risk of genomic integration e Up to 10X hig
7. of the lon Torrent platform combined with key workflow advantages designed specifically for regulated laboratory environments and in vitro diagnostic applications Used in conjunction with lon PGM Dx library kits lon OneTouch Dx template kits lon PGM Dx sequencing kits lon 318 Dx chip kits and data analysis software the lon POM Dx Platform unu nb s is intended for targeted sequencing of human genomic DNA from peripheral whole blood samples Learn more at lifetechnologies com pgm dx Enhance your template prep and lon Chip loading lon Chef System The lon Chef System simplifies the workflow for the lon Proton and lon POM Systems providing a fully automated high throughput solution for reproducible template preparation and chip loading Simplified workflow Reducted variability Helps save time and labor Simplifies and automates Enables reproducible chip Delivers walk away automation template preparation and chip loading for users of any with typically just 15 minutes of loading in a single system experience level setup time Delivers easy setup and Provides sample and reagent Helps increase your lab s operation with intuitive touch tracking with onboard tracking productivity with the ability to screen interface system perform overnight runs Find out more at lifetechnologies com ionchef TM TM lon PGM Hi Q Sequencing Kit Offers rapid accurate and cost effective sequencing for you
8. studies GWAS requires validation using a large number of samples We re making it simpler by adding 1 7 million new TaqMan SNP Genotyping Assays increasing our collection total to 6 2 million And like all of our TaqMan Assays the performance of these new genotyping assays is backed by the TaqMan Assays Guarantee Spend time on results not assay design or optimization Search for assays now at lifetechnologies com taqmansnp life in the lab Wow For researchers working with mammalian model systems recombinant proteins formed by mammalian expression systems are more likely to have the expected posttranslational modifications to fold correctly and to function properly in those models Sino Biological uses mammalian expression systems to generate recombinant proteins that are as close to the native form as can currently be produced Life Technologies through a partnership with Sino Biological brings you an expanded portfolio of recombinant proteins for a variety of different applications A d Proteins priced below competitive brands Mammalian expressed proteins with gt 90 purity Proteins with different Sizes species and tags meeting different needs We offer wide selection of recombinant proteins Human Canine Mouse Dog Rabbit Influenza Rat Monkey Aequorea Victoria RSV Mammalian E coli Insect Oncology Inflammation Immunology Angiogenesis Stem cell research Drug Development Neurob
9. Almost all lives are at some point touched by cancer Fundamentally changing how this disease is diagnosed and treated will make life better for all 6 6 6 0 6 06 9 06 06 9 9 0 0 9 0 0 0 0 9 0 O0 9 6 0 9 06 O 0 0 GO 9 O0 O 9 0 0 0 6 O0 0 0 GO 0 6 0 O 0 06 O0 0 0 GO 0 9 0 0 9 0 O0 0 0 O0 0 0 O0 0 6 0 O 9 6 O 0 0 O 0 9 0 O 9 0 O0 0 06 GO 0 9 0 O 0 09 O 9 0 O 0 0 0 0 0 O 9 9 O0 6 O0 0 9 06 9 9 GO 9 6 6 0 9 6 6 9 6 6 lon AmpliSeq BRCA1 and BRCA2 Panel A community panel designed with leading researchers It contains primer pairs that target the coding regions of the tumor suppressor genes BRCA1 and BRCAA2 which have been implicated in hereditary breast and ovarian cancers e High coverage of the target region e High performance amplicons and uniformity e Fast turnaround time coupled with throughput flexibility e Adoptable by other translational research labs Learn more at lifetechnologies com ampliseqcommunity The lon AmpliSeq BRCA panel enabled us to develop a quick and easy high throughput workflow with minimal hands on time It is an accurate solution of the 65 samples tested to date we detected all expected mutations Dr Arjen Mensenkamp Clinical molecular geneticist ufetechnologies com IMMUNOPRECIPITATION IS GOING hile Sepharose beads are still great for purification Why is the number of of large amounts of protein or antibody it s not well immunoprecipitation IP sui
10. Conventional cancer therapies fail in many patients and for women with triple negative breast cancer the picture Is particularly grim But a new study employing a personalized sequencing based approach Is bringing new hope to women who a short time ago had little reason for optimism Research was conducted using retrospective samples from 13 women with triple negative breast cancer who had previously failed all other treatments These samples were sequenced on Life Technologies instruments at the Translational Genomics Research Institute 6 0 6 9 6 6 6 9 0 0 0 9 0 6 0 9 0 O0 9 0 0 O 0 9 0 0 0 9 O 0 9 0 GO 9 09 0 0 0 9 0 0 0 0 0 O0 9 0 O 0 9 9 GO 0 GO 9 0 O 0 0 GO 9 9 6 0 9 9 GO 9 0 0 O 9 0 0 O 9 6 0 9 9 0 9 9 6 o 9 9 6 6 Doctors examined the DNA sequence obtained from tumor tissue and placed each woman on therapies selected by taking the sequencing data into consideration Some women were enrolled in clinical trials for new drugs not yet on the market 0609206206202020606206292209020206202020602029029020202060206020202062929029029020629920206020202020296029020202060292090290206029202209002902920290292020292209029029029229209206020290290292902929290292960202909292902920292992929929299929299 We are collaborating on a global basis with academic and pharmaceutical partners to show how sequencing may help solve what is one of the largest and most complex medical problems affecting our world
11. DEVICE Obtain greater sensitivity and reproducibility as compared to manual processing methods Automated hands free processing enables better blot to blot consistency while using up to 80 less primary antibody View robust data at lifetechnologies com ibind Dynabeads Protein G are so easy to work with and the background is minimal don t know what would ever do without them now FAST AND INTUITIVE PowerEase Power Supply Designed specifically for mini gel electrophoresis and western transfer PowerEase Power Supply from 3 699 love the ease of this reagent PS0091 PowerEase 90W Power Supply Just 3 699 Faster and cleaner PS0301 PowerEase 300W Power Supply Just 6 399 Use promotion code P1252760 Offer expires March 13 7015 lerms and conditions apply Find out more at lifetechnologies com powersupplies Get a Mini Gel Tank or Mini Blot Transfer Modules free Buy 5 boxes of selected protein mini gels in any combination at list price and receive a Mini Gel Tank free Buy 7 boxes of selected protein mini gels in any combination at list price and receive two Mini Blot Transfer Modules free Use promotion code RHKPROA Please contact your sales representative when you place order Offer expires March 13 2015 Terms and conditions apply Find out more at lifetechnologies com lifeinthelabhk12 litetecnnologies com 23 ANTON MBPS E OM THE CULTURE DISH TO A VIDEO SCREEN GIBCO
12. Do you have a favorite list of genes you would like to target with a collection of siRNAs or miRNAs Would you like an expert bioinformatics team to identify a gene list covering your desired disease or pathway We can prepare a custom library against any set of human or mouse genes For more information and quote requests go to lifetechnologies com sirna and lifetechnologies com mirnalibraries N D I md D Q D D m 3 Q TAKE YOUR LAB WALLS FROM DRAB TO FAB Two new Invitrogen posters available Showcase your area of interest with the latest installments in our genome editing poster series Choose Genome editing with engineered nucleases or The evolution of genome engineering Request your free poster at lifetechnologies com genomeeditposters ALL AROUND FREEDOM UNLIMITED GENOME TARGETING GeneArt PerfectMatch TALs provide custom DNA binding proteins for accurate DNA targeting and precise genome editing Unlike other technologies and traditional TALs that limit the choice of targets or provide ambiguous results GeneArt PerfectMatch TALs enable the targeting of any locus in the genome Experience the power of GeneArt PerfectMatch TALs lifetechnologies com tal EASILY DETERMINE THE CLEAVAGE EFFICIENCY OF CRISPR TAL The GeneArt Genomic Cleavage Detection Kit allows rapid and reproducibly determine the cleavage efficiency of your CRISPR and TALs e Easy no genomic DNA isolation direc
13. Kit provides optimal permeabilization conditions for most cell types Request a free sample at lifetechnologies com freefixedsamples litetecnnologies com SISA eue TS Senior Research Fellow Institute of Medical Biology Singapore Role of microRNAs in cutaneous wound healing in human skin We are studying how microRNAs regulate wound healing using a human ex vivo organ culture system and cultured keratinocytes as tools In order to study the function of a specific miRNA or its target mRNA it is essential to manipulate their levels The chal lenge is the intracellular delivery of miRNA mimics inhibitors siRNAs at close to 10096 efficiency in order to see the effect on cells Keratinocytes are known to be difficult to transfect After trying multiple transfection reagents and vendors we tried Lipofectamine RNAIMAX Transfection Reagent Most other transfection reagents give about 50 7596 transfection efficiency Lipofectamine RNAIMAX yields 90 transfection efficiency with siRNAs and was able to achieve more than 80 knockdown of my target genes On top of that Lipofectamine RNAIMAX also works well for miRNA mimic inhibitor transfection Toxicity of transfection reagents to keratinocytes which encountered with other transfection reagents was minimal with Lipofectamine RNAIMAX and have transfected cells multiple times without adverse side effects Bleaching of fluorescent signals in confocal imaging of cell
14. PRODUCTS INFORMATION AND SCIENTAINM NT ISSUE 12 WINTER 2015 amp b Science Advance Ee That Will Change fing page 16 Doing Genome Editing ae Use Our Handy Ma buc MN D i page 18 j Kees a Disease Spotlight Breast Cancer v page 21 What s Your PCR 10 page 20 Better Together Thermo Fisher scientific Integration Update page 3 Reagent Rewards App How Much Will You Save page 31 Invitrogen Applied Biosystems 0 Molecular Probes ax Ambior lon Torrent Buy 1 get 1 free Free magnet for Just 3 699 for se 3p on SuperScript every purchase PowerEase VILO amp of Dynabeads Bt en 90W Power pages i immunoprecipitation Supply E product T page 22 ninh a i 4 knw ir Fir ums em E FL dein SIMPLY STUNNING FLUORESCENCE IMAGING HAS NEVER BEEN EASIER Innovative EVOS cell Invitrogen Countess ReadyProbes More than 40 000 imaging systems FL Automated ready to use imaging primary antibodies Minimizes the complexities of Cell Counter reagents Offers a wide variety of primary microscopy without compromising Everything counts but nothing counts No Calculations antibodies against key signaling performance like the Countess Il FL Automated No Dilutions proteins cellular markers lifetechnologies com evos Cell Counter No Resuspensions and organelles as well as cell lifetechnologies com countessll lifetechnologies com readyprobes junct
15. accurate optimization Request a demo at Find out more at ufetechnologies com REAGENT REWARD TagMan Flexible Content Panels Looking at a large set of genes in a pathway disease or biological process can be a great way to identify markers of interest for a given phenotype TaqMan flexible content panels offer an easy way to select and design a combination of assays in the format that matches your experiment Panels at launch Gene Expression flexible content panels Alzheimer s Disease Glycosylation Angiogenesis Hepatotoxicity Apoptosis Hypertension Cardiotoxicity Inflammation Cardiovascular Nephrotoxicity Cell Cycle p53 Signaling Diabetes Parkinson s Disease Endogenous Controls SNP Genotyping flexible content panels Alzheimer s Disease Pharmacogenomics Autism Type 2 Diabetes Experience the convenience of predesigned panels with the flexibility to modify the content in a few clicks B rcgwss cards oler format Bdk 4 confirm Drder Erie m gr c3 pr T gh a5 4 Zn ac Pr mm ort Hr he d ey Pe ir Cw EE DS espe ii EF ps Eod LUE WD Le parma biu med och cn TNTE IEEE avo rik c updmim pour irre Pre ary Pee Seed eae re Fie ual to ee ae fee fue eee Try it now at lifetechnologies com flexiblepanels An assay for every genetic occasion 6 2 million TaqMan human SNP genotyping assays are now available for GWAS validation Identification of potential causal variants by genome wide association
16. d are working on that we think will change everything EXOSOMES Cells talk all day their social network of choice is a cell to cell messenger service known as exosomes Like Snapchat photos the messages passed via exosomes shouldn t always be sent Scientists are working on ways to Isolate these messengers stop the bad messages i e spreading of disease and program in positive messages i e deliver drugs to cure disease GENOME EDITING Bad coding happens Your DNA made up of 4 building blocks A T C and G may take a wrong turn along the path of life To set the code back on course scientists in the labs around the world are working on editing technologies i e CRISPRs and TALs to fix broken code reverse detrimental mutations INDUCED PLURIPOTENT STEM CELLS IPSC Embryonic stem cells have come a long way since 1998 In 2006 scientists showed how existing cells such as skin cells could be reprogrammed into stem cells and re differentiated to form a different type of cell in the body such as a neuron Imagine Parkinson s disease patients being healed with their own skin cells but don t just imagine clinical research trials could begin soon life in the lab rce initiatives to advance personalized FOOD SAFETY Remember the 2011 epidemic in Germany sickened nearly 4 000 people Food borne illness hits 1 in 6 Americans Ni Industry government and academic scien
17. eck microspheres are provided in suspension or mounted on slides and in several different multicolored configurations of fluorescent ring stains with or without contrasting fluorescent dyes throughout the bead The sharp ring stains produce a striking visual representation of instrument misalignment or other aberrations making them ideal reference standards Selected Products Catalog Name Size F24633 FocalCheck Fluorescent Microspheres Kit 6 um mounted on slides 1 kit F7238 FocalCheck Microspheres 15 um fluorescent green ring stain dark red throughout 0 5mL F36909 FocalCheck Fluorescence Microscope Test Slide 1 for alignment intensity amp calibration 1 each A b 7 us Figure 1 Confocal laser scanning microscope optical cross sectioning and alignment with FocalCheck microspheres ms c A Serial optical sectioning from top to bottom along the z axis of ring stained microspheres reveals a continuous pattern of disc to ring to disc images Ore B The diameter of the fluorescent ring or disc seen is dependent on the depth of the optical focal plane C Inthe confocal laser scanning microscope separate light paths exist i a for UV and visible wavelengths and emitted fluorescence is detected by different photomultipliers Migrant C Proper optical alignment in three dimensions may be obtained with either of two types of FocalCheck microspheres 1 microspheres with a ring stain and a second
18. extraction after RNAlater stabilization solution to your samples for fast more than 25 years referenced in over 100 000 scientific RNA protection publications lifetechnologies com rnalater lifetechnologies com trizol WHAT S LURKING ON YOUR BENCHTOP No one ever said that working with RNA would be easy RNase lurks around every corner like a gremlin But by following basic good laboratory practices RNA degradation need not be an issue Our Ambion RNA experts have compiled some tips to help repel an RNase attack Including e Preparing your workspace e Sample harvesting e RNA purification e RNA storage Check out the helpful tips at lifetechnologies com rnase life in the lab daid ajdwes Q How can inactivate DNase I ONE STEP FORWARD Meet the fast 1 step kits for real time PCR analysis Add 1 uL of a 25 mM EDTA solution to a 10 uL reaction mixture containing 1 unit DNase Amplification Grade or at a 1 1 molar ratio of Mg EDTA to chelate the Mg ions in the DNase buffer Heat for 10 min at 65 C Note It is vital that the EDTA be added Caina concenti auon ol ai lea ci Sound too good to be true mM prior to heat inactivation to avoid Mg dependent RNA hydrolysis Ambion Cells to C kits offer expression analysis directly from cultured cells without RNA purification And the 1 step RT PCR master mix allows all reactions to be performed in a single tube Check out the video and see a demon
19. fetechnologies com WE RE HERE TO HELP YOU SAVE To make your research a bit more affordable we ve featured many money saving promotions throughout this issue For your convenience some favorites are listed here If you d like more information about any of these promotions or other promotions found in this issue simply go to lifetechnologies com lifeinthelabhk12 P a On selected agarose Free Gibco Life of Cells minifigures On selected SuperScript VILO Use promotion code MHAGCT With qualifying Gibco cell culture purchase cDNA Synthesis Kit and Master Mix No promotion code needed Use promotion code MHAGCT i wrp Save 30 on selected PCR Plastics Purchase a Dynabeads immunoprecipitation Purchase Lipofectamine Use promotion code P1252760 product and get a free magnet or 50 off for MessengerMAX reagent and DynaMag 2 Magnet receive a cool T shirt Use promotion code RHKCATA for free magnet and No promotion code needed RHKDYNA for DynaMag 2 Magnet Please contact your sales representative when you place order Free tank transfer modules Buy 5 boxes of selected protein mini gels in any combination at list price and receive a Mini Gel Tank free Buy 7 boxes of selected protein mini gels in any combination at list price and receive two Mini Blot Transfer Modules free Use promotion code RHKPROA Please contact your sales representative when you place order Offer expires March 13 2015 Terms and co
20. fication of NGS libraries with digital PCR lowers sequencing workflow costs Next generation sequencing NGS libraries can be quantified with minimal sample handling and without the need to generate a standard curve using digital PCR This method enables accurate and precise library quantification a critical step in both the lon Torrent and lllumina workflows allowing for maximizing sequencing yields downstream To achieve this high degree of precision a TagMan Assay designed to span both the forward and reverse adapters specific to each library is available ufe in the lab PubGo app Access product information from papers with a single click Want to quickly identify relevant products and related material from a journal article With the PubGo app you can Do you have an older paper that cites products that are no longer available Not a problem The PubGo app helps find replacement products or suggest possible alternatives Find out more and download the app at lifetechnologies com pubgo Monkey mouse fish frog pig oh my Working with these or other animals Don t lose your precious time in design when we have the solution you need Whatever the animal model you study we have a TagMan Gene Expression Assay ready for you Find out more at lifetechnologies com taqmangeneexpression Primer pairs when you need them Primer Designer Tool is a free online tool that helps you find the PCR Sanger primer
21. fluorescent stain throughout the bead or Orge Utm 2 microspheres containing three different fluorescent ring stains Adjust Color Registration With Tetraspeck TetraSpeck Fluorescent Microspheres facilitate the chromatic registration of conventional fluorescence microscopes confocal laser scanning microscopes and associated image processing equipment They are available in five diameters spanning the range from subresolution to nearly cell size particles They are stained throughout with four different fluorescent dyes yielding beads that each display four well separated excitation emission peaks Selected Products Catalog Name Size T7279 TetraSpeck Microspheres 0 1 um fluorescent blue green orange dark red 0 5mL T7280 TetraSpeck Microspheres 0 2 um fluorescent blue green orange dark red 0 5mL Learn more at lifetechnologies com microscopecalibration TURN YOUR REAGENTS INTO REWARDS Have you downloaded the Reagent Rewards app When you join the Reagent Rewards program you ll earn points each time you purchase and scan a qualifying product on your smart device We ve added more qualifying products to the program so now you can build up those points faster than ever and redeem them for gifts and discounts Reagent Rewards members can earn points when they buy qualifying e Endpoint PCR reagents e Real time PCR reagents e Cloning and competent cells e Sample preparation reagents e GeneArt Gene S
22. g gene expression analysis and profiling Find in depth technical information on SuperScript and SYBR reagents from peer reviewed publications for your real time PCR experiments Check out what your fellow researchers are publishing lifetechnologies com superscript sybr citations SuperScript VILO Master Mix For improved qRT PCR assay dynamic range Optimized for reliable generation of first strand cDNA the SuperScript VILO cDNA Synthesis Kit and SuperScript VILO Master Mix are ideal choices for synthesizing cDNA for qRT PCR The formulations are designed for enhanced cDNA synthesis efficiency and can be used with 1 pg to 2 5 ug of input RNA in a 20 uL reaction For additional convenience SuperScript VILO Master Mix features a single tube format with a simplified workflow Find out more at lifetechnologies com vilo REAGENT On selected SuperScript VILO cDNA Synthesis Kit and Master Mix Use promotion code MHAGC1 Offer expires March 13 2015 Terms and conditions apply Find out more at lifetechnologies com lifeinthelabhk12 REWARD ufe in the lab ELEGANT DESIGN SMART CHOICE SimpliAmp Thermal Cycler The SimpliAmp Thermal Cycler is an easy to use accurate and small thermal cycler designed to fit any lab s needs for everyday PCR It is designed with reliable Applied Biosystems engineering including innovations like the responsive color touch screen and VeriFlex technology for simple and
23. her cleavage efficiency with CRISPR mRNA lifetechnologies com messengermax No promotion code needed Offer expires March 31 2015 Terms and conditions apply Find out more at lifetechnologies com messengermax GET BEAUTIFUL RESULTS AGAIN AND AGAIN WITH INSTRUMENTS POWERED BY MOLECULAR PROBES REAGENTS BOUNDARY BREAKING Attune NxT Acoustic Focusing Cytometer The Attune NxT Acoustic Focusing Cytometer combines superior precision speed and sensitivity in a modular design that can grow with you With up to 4 lasers and 14 colors the Attune NxT cytometer is designed to accommodate most experimental design needs and lab budgets Boundaries beware lifetechnologies com attune FLOW CYTOMETRY REAGENTS LIVE DEAD fixable dead cell stain kits Now you have the flexibility you need choose your color laser and pack size lifetechnologies com livedeadfixable BackDrop Background Suppressor for Microplate Assays Designed to effectively suppress background fluorescence In live cell microplate assays New When this reagent is used with green fluorescent dyes or proteins Luhr L the signal to noise ratio improves greatly O A E Er Check out the new products at lifetechnologies com newprobes FLOW CYTOMETRY RESOURCES Answers to your flow cytometry questions Get your free copy of the new Molecular Probes Flow Cytometry Products and Resource Guide is lifetechnologies com flowguide eee
24. iology Microbiology Signal transduction Virology Apoptosis Metabolism Shop now at ufetechnologies com E LOAD AND 60 Tracklt DNA Ladders Our most convenient solution for estimating molecular weight and DNA mass There is no need to heat mix or dilute these ladders prior to loading them on your gel lifetechnologies com ladders life in the lab I CHOOSE How about reliability and innovation mm IL DNA sabe Se FAST AND EASY TOPO cloning kits TOPO cloning kits use a o9 minute room temperature reaction that requires no gel purification or post PCR modifications and yields up to 99 recombinants lifetechnologies com topo VENT HEAT ADD STAIN POUR Introducing a simplified solution to agarose gel preparation ReadyPouch Agarose Gels are packaged In a ready to microwave pouch that contains the right amount of agarose and buffer for a 100 mL gel They come with an easy to use stain dropper All you need to do is vent the pouch heat in a microwave oven add stain and pour lifetechnologies com readypouch On selected agarose Use promotion code MHAGC1 Offer expires March 13 2015 Terms and conditions apply Find out more at lifetechnologies com lifeinthelabhk12 9 MINUTE LIGATION ExpressLink T4 DNA Ligase Very fast 5 minute ligation of blunt or cohesive end DNA fragments or PCR fragments with A overhangs in 15 minutes at room temperature 25 C lifetech
25. ions and CD markers lifetechnologies com antibodies life in the lab A LETTER FROM MARK STEVENSON EXECUTIVE VICE PRESIDENT AND PRESIDENT LIFE SCIENCES SOLUTIONS THERMO FISHER SCIENTIFIC As one company we are creating new opportunities to support the convergence of life sciences tools and diagnostics to improve patient care now and in the future ufetechnologies com Dear Valued Customers By now you are certainly aware that Thermo Fisher Scientific acquired Life Technologies in February 2014 Our goal through the combination is to bring the best of both companies together to create unrivaled value for our customers working in research life sciences specialty diagnostics and applied markets Our combined offering gives customers access to an even broader base of cutting edge technologies such as genomics and proteomics to accelerate life sciences research and improve human health Our complementary strengths in biosciences and bioprocessing accelerate drug discovery development and production And as one company we are creating new opportunities to support the convergence of life sciences tools and diagnostics to improve patient care now and in the future As a part of Thermo Fisher Scientific the world leader in serving science we are In an even stronger position to help our customers achieve their goals for innovation and productivity Our 50 000 employees are proud to work with you to fulfill our
26. luding optimized procedures and products that will help you get the consistency your lab work demands Students completing this course will be eligible to take the free Gibco Cell Culture Basics certification test and earn the corresponding certificate Learn more www canvas net courses cell culture basics life in the lab TAKE CARE OF YOUR CELLS Gibco media supplements cells and cell culture reagents are designed to deliver reproducibility and performance for results you can count on every day lifetechnologies com gibco Mr Jurkat l m Mr Jurkat from the immortalized cell line of the late 1970s Maybe you already use me in your T cell experiments or cancer research or perhaps you know me for my high IL 2 production It s not bragging If it s true litetecnnologies com Ms HUVEC Mr Jurkat Stem Cell Vero CHO S Feb 16 Mar 6 Mar 23 Apr 3 Jan 19 30 Feb 2 13 Mar 9 20 2015 2015 2015 2015 2015 How to get the Life of Cells minifigures Now you can start collecting Gibco Life of Cells minifigures based on our famous Gibco Life of Cells video series characters Simply collect ten cap labels from Gibco media and reagents stick them on the Gibco scoreboard and return the Gibco scoreboard to us on specified dates It s that easy Be one of the first researchers to get your hands on the limited edition Gibco Life of Cells minifigures Find out more at lifetechnologies com life
27. mission which is to enable our customers to make the world healthier cleaner and safer Sincerely MPE Mark Stevenson FAST AND AFFORDABLE gene expression analysis with lon lorrent solutions The lon Proton System and lon AmpliSeq Transcriptome Human Gene Expression Kit ast and affordable gene expression analysis from limited as little as 10 ng of total RNA or degraded samples such as FFPE tissues is now a reality With simultaneous amplification of more than 20 000 RefSeq genes in a single tube the lon Proton System and lon AmpliSeq Transcriptome Human Gene Expression Kit transform translational and cancer research in tumor profiling and biomarker discovery lifetechnologies com amplisegtranscriptome See the data comparing FFPE and fresh frozen samples under Datasets tab lifetechnologies com ioncommunity More differentially expressed genes detected than microarrays The Venn diagram shows the concordance of significant differentially expressed genes DEGs between MAQC microarray and lon AmpliSeq Transcriptome Human Gene Expression Kit data hd l Eom y r Affy Hu 133A 2 microarrays Detection of differentially expressed genes DE6s by using the lon AmpliSeq Transcriptome Human Gene Expression Kit ufe in the lab lon POM Dx System Empowering diagnostic assay development with next generation sequencing The lon PGM Dx Instrument System delivers the speed and simplicity
28. nditions apply Join the Reagent Rewards program to earn points each time you purchase and scan qualifying products Look for this icon on qualifying products e Endpoint PCR reagents ae T e Real time PCR reagents Aperi e Cloning and competent cells e Sample preparation reagents Just 3 699 for PowerEase 90W Power Supply e GeneArt Gene Synthesis reagents Just 6 399 for PowerEase 300W Power Supply e Molecular Probes fluorescence reagents Use promotion code P1252760 Offer expires March 13 2015 Terms and conditions apply lifetechnologies com reagentrewards Invitrogen Applied Biosystems Gibco Molecular Probes Novex lon Torrent For Research Use Only Not for use in diagnostic procedures Abbreviated terms and conditions Use promotion codes as noted All promotions are open to qualified academic end users in Hong Kong Offers will apply to qualifying orders received by Life Technologies no later than March 13 2015 unless otherwise noted or until promotional supplies are depleted whichever comes first Quantities may be limited and other restrictions may apply to these promotions Promotion code must be presented at the time of order Discounts apply to the list price in effect at the time the order is received by Life Technologies Each promotion cannot be combined with other discounts or promotions For Dynabeads immunoprecipitation kit promotion customer can use the offer only once Offer only a
29. nologies com expresslink REAGENT REWARD CONVENIENCE Then Invitrogen instruments and reagents are the clear choice I md D a I mi QD um 2 T O Q lt QUANTITATION ISN T THE SAME WITHOUT IT Sensitive accurate and affordable RNA DNA MicroRNA t ET IT and protein quantitation The Qubit 3 0 Fluorometer enables accurate data in a few seconds LOVE IL with touch screen controlled operation and your choice of assays Why do you need the Qubit 3 0 Fluorometer e t quickly and accurately quantifies dsDNA ssDNA RNA MicroRNA and protein in 5 seconds per sample e t uses as little as 1 uL of sample e t stores up to 1 000 sample results e t features a large 5 7 inch color touch screen for easy workflow navigation e t offers the ability to personalize your Qubit fluorometer with the assays you run most add new assays or even create your own with the MyQubit design tool Do you detect d glucose or glucose oxidase Amplex Red Glucose Glucose Oxidase Assay Kit detects d glucose or glucose oxidase on Qubit 3 0 Fluorometer e Detect concentrations as low as 3 uM d glucose or 0 05 mU mL of glucose oxidase e Format allows for multiple time point measurements e Designed for minimal autofluorescence interference Do you measures free cholesterol and cholesteryl esters Amplex Red Cholesterol Assay Kit provides a sensitive rapid and simple fluorometric method for detecting very
30. ofcells The Life of Cells minifigures Where will yours go Tweet us your picture dGibco with lifeofcells Or post on our G ibco Facebook page facebook com gibco YOUR STEM CELLS THRIVE WITH THE ESSENTIALS A seamless system for feeder free xeno free culture When used together the Essentials culture media streamline the generation expansion and differentiation of pluripotent stem cells PSCs by consistently maintaining all three steps in a feeder free xeno free system feeder free xeno free system Find out at lifetechnologies com essentialmedia COMPONENTS 8 IS GREAT Gibco Essential 8 Medium is a feeder free xeno free medium originally developed in the laboratory of stem cell research pioneer James Thomson which contains only the 8 essential components needed to grow and expand pluripotent stem cells Why are 8 media components better than 20 Essential 6 Medium Used with Essential 8 Medium enables more efficient cell reprogramming My experience with Essential 6 Medium is very positive iPS colonies are bigger with more defined colony borders compared to our standard induction Essential 8 Medium Consistently grow and expand pluripotent stem cells while minimizing variability Timo Otonkoski Biomedicum Stem Cell Center Finland Essential 8 Medium does exactly what you need it to do And because it s so simple it can be made in a more controlled way Emile Nuwaysir PhD
31. pplies to a maximum of one DynaMag 2 magnet cat no 12321D at 50 discount or one free magnet Cat no 10916S per customer By placing an order you warrant that you are not prohibited by employment contract or law from accepting a gift from Life Technologies All offers are void where prohibited by federal state provincial or local laws or regulation or agency institutional policy Other restrictions apply Life Technologies Limited reserves at its sole discretion to determine the eligibility to receive the special offer To see complete terms and conditions go to lifetechnologies com lifeinthelabhk12
32. r research The lon POM Hi Q Sequencing Kit contains reagents and consumables for robust and highly accurate sequencing of 200 and 400 base pair libraries using the lon OneTouch 2 System combined with the lon PGM System e Robust sequencing for lonAmpliSeg panels improved indel sequencing accuracy observed across targeted resequencing panels especially with longer amplicon designs e Higher quality de novo assemblies up to 90 decrease in indel error rates for microbial sequencing e Unified workflow one sequencing kit to support all current lonPGM sequencing research applications Find out more at lifetechnologies com iontorrent lifetechnologies com QUANTSTUDIO 3D DIGITAL PCR RARE MUTATION SOLUTION APPLIED BIOSYSTEMS INSTRUMENTS AND REAGENTS IN r gig Guanes a Quantify rare mutations at a rate as low as 0 1 with digital PCR Need a highly sensitive and precise solution for the confirmation or characterization of rare mutations Then you need TagMan SNP Genotyping Assays These assays have been wet lab validated and optimized on the QuantStudio 3D Digital PCR System to identify key somatic mutations in these cancer genes KRAS BRAF EGFR PIK3CA and JAK2 Find the wet lab validated digital PCR assays at lifetechnologies com dpcr raremutation See the QuantStudio 3D Digital PCR System at lifetechnologies com quantstudio3d Next Generation Sequencing NGS Library Quantification Absolute quanti
33. s tissues after immunocyto histochemistry was a major obstacle in my research Shifting to Molecular Probes fluorescent secondary antibodies with Alexa Fluor conjugates instead of conventional FITC TRITC dyes gives much more stable fluorescent signals without significant loss due to bleaching Consistent target MRNA knockdown or miRNA modulation was achieved using Lipofectamine RNAIMAX with 100 confidence Plus stable knockdown of target genes was seen up to 5 days posttransfection Success n N P o c w o life in the lab OBTAIN THE HIGHEST QUALITY DAT YOUR SYSTEM CAN DELIVER FLUORESCENT MICROSPHERES FOR CALIBRATING MICROSCOPES Regular instrument calibration and adjustment are required for high precision imaging of fluorescent probes 1 particularly in multicolor applications such as confocal laser scanning microscopy that involve multiple exposures repetitive scans or three dimensional sectioning We offer a range Molecular Probes calibration products to help you achieve optimum performance from your imaging system Confirm Color Alignment With Focalcheck Beads Our Molecular Probes FocalCheck fluorescent microspheres are specifically designed for examining the alignment sensitivity and stability of confocal laser scanning microscopes They are particularly useful for confirming the optical sectioning thickness Z resolution in three dimensional imaging applications FocalCh
34. stration of the simplified protocol DNases in DNA free and Turbo DIN ees kte can be nmacivated with the included DNase Inactivation Reagent Find out more lifetechnologies com cellstoct WHY STUDY SINGLE CELLS Because tissues are composed of heterogeneous mixtures of cells gene expression measurements based on the homogenized population don t account for the small but critical changes occurring in individual cells Single cell analysis can be critical in applications such as candidate drug screening cell differentiation and stem cell studies and measuring individual cell responses to specific stimuli Ambion Single Cell to CT qRT PCR Kit e Pre optimized workflow for real time RT PCR from single cells e Maximum sensitivity for single cell analysis e Complete kit convenience and easy to follow protocol Ambion Single Cell Lysis Kit e Enables RNA preparation for 1 10 cells e No sample loss ensures maximal sensitivity e Compatible with common enzymatic reactions Find out more at lifetechnologies com singlecell ufetechnologies com b SCIENCE ADVANCES THAT WILL CHANGE EVERYTHING We live in a century of amazing science never before have scientists been so empowered with access to advanced technologies global connectedness and M i medicine or prevent food borne illnesses Healthier cle ier Safer that s the world we all are working for Here we highlight 5 breakthroughs research labs across the worl
35. t PCR amplification e Rapid 4 hour total processing time e Quantative gel band density directly correlated to target indel introduction Find out more at lifetechnologies com genomiccleavage ufetechnologies com WHAT S YOUR PCR T hink you know everything there is to know about PCR lest your knowledge with our fun quiz Answers are shown below but remember no peeking 1 In 1989 the journal Science named this enzyme as its first Molecule of the Year EJ Restriction endonuclease IJ Tag polymerase I33Methylase F Reverse transcriptase 2 PCR entered the mainstream media via an episode of this television show EJ A Different World J Star Trek The Next Generation I33 Unsolved Mysteries FF Murder She Wrote life in the lab 3 DNA fingerprinting using PCR was accepted in court as reliable forensic evidence and became well known in the US when prosecutors relied on DNA evidence in this person s criminal trial EJ Naomi Smith IJ O J Simpson p FA Kirk Bloodsworth 4 A basic PCR reaction has three phases exponential linear and plateau Which one of these three PCR techniques measures the amount of accumulated PCR products EJ Digital PCR J Real time PCR IA Endpoint PCR 5 In 1983 Kary Mullis had a Eureka moment that led to the discovery of PCR while doing what EJ Driving a winding road J Drinking a beer at a pub FA Walking through an art exhibition
36. ted for the smaller scale isolation of specific publications citing proteins and protein complexes more common today Dynabeads magnetic mM ME beads skyrocketing The key today is balancing the capacity yield reproducibility o purity and cost e Minimal nonspecific binding oA Dynabeads magnetic E Iyraceadr magnete beats e Easy handling d a EM Separation is the fastest Ae ain peel ilm growing method for IP and i 0 NB Apu no centrifugation ChIP This graph shows the n percent increase decrease of a e Allows you a 1 published papers using top IP 14 i and ChIP products Source sind o Goode Schola Oct 2014 ep i T omn d reproducibility Ten purity and cost J PALALANAL ID MAV TII C C AFDITA IKEN amp L U ld B E Ww Y id i J U l J b Y Y LY L U Check out our myth busting video series at MYTH FACT Almost all background is removed using Dynabeads magnetic beads because all antibodies are accessible on the smooth bead surface Background can t be avoided limiting nonspecific background Pre clearing is necessary Pre clearing step is unnecessary with Dynabeads magnetic beads You can save time and you use half the amount of solid phase to get good IP results which helps save money The high capacity of Sepharose beads comes from high surface area that may also traps a lot of unwanted protein and wastes a lot of Se a al antibody Even with lo
37. ti re working together to 5 sequence nearly 200 species of E coli to identify toxic strains and aid development of new tests that may help prevent future food borne outbreaks Breakthroughs in this field are continuous and may help prevent contaminations that are Z j In c S im L human and animal food in the future GENETIC ANALYSIS AND BIOINFORMATICS You ve heard of genome sequencing you may have even done the 23andMe genotyping test With the dual advancement of genome sequencing nem and supercomputers bioinformatics analysis of genetic information is impacting areas Mn disease research and drug discovery to human identification One field of special note pharmacogenomics the study of how a person s genes affect their response to drugs is bringing personalized medicine to universal reality LI winter 2015 A Thermo Fisher Scientific Brand ufetechnologies com LOOKING FOR INVITROGEN GENOME EDITING PRODUCTS START HERE miu DESIRED GENE EDIT Are you working Ma um ELL Yeu Ue tied mm ceils for cammercul ia applications Do you have target sequence constraints Ge Lack of PAM sequencel Learn more here lifetechnologies com geneedit Wha Is Your doanstream apelication or hast cell type For donor plasmid and oligo DNA synthesis contact geneartsupport dlifetechnologies com life in the lab ee CUSTOM RNAi LIBRARIES THE ULTIMATE IN FLEXIBILITY
38. ts of washing you will end up with unwanted background Good capacity with high yield is best Dynabeads magnetic beads are expensive With no pre clearing and less antibodies used Dynabeads magnetic beads help save you money by balancing optimal capacity yield reproducibility and purity b e Use promotion code RHKCATA for free magnet and RHKDYNA for DynaMag 2 Magnet Please contact your sales representative when you place order Offer expires March 13 2015 Terms and conditions apply d Customer can use the offer only once Offer only applies to a maximum of one DynaMag 2 magnet cat no 12321D at 50 discount or one free magnet Cat no 109165 per customer da Find out more at life in the lab Protein research needs have changed since the introduction of Sepharose beads in the 1970s ONE TANK 181 GELS MINI GEL TANK Now run your favorite NUPAGE Novex and Bolt gels in the same tank Our Mini Gel Tank enables versatility easy Loading and enhanced visualization Explore more choices at lifetechnologies com minigeltank A6o o1g ula o1d What would it take for you to change your 4 protocol for IP e Easy workflow e Consistent performance e Ability to remove almost all background e Faster protocol time No problem Dynabeads magnetic beads can do all of the above and more Here s what people are saying about the kits SUPERIOR WESTERNS AND ANTIBODY SAVINGS IBIND WESTERN
39. ynthesis reagents e Molecular Probes fluorescence reagents So what are you waiting for Find out more at lifetechnologies com reagentrewards REAGENT Look for this icon throughout the issue REWARD for qualifying products Available for iOS and Android devices Terms and conditions apply Life in the Lab Issue 12 Winter 2015 Life in the Lab is published quarterly by Thermo Fisher Scientific e 5791 Van Allen Way Carlsbad CA 92008 For Research Use Only Not for use in diagnostic procedures Life Technologies Limited Units 01 09 13 amp 15 18 Level 13 Tower 1 Kowloon Commerce Centre No 51 Kwai Cheong Road Kwai Chung N T Hong Kong General Line 31077600 Customer Care Hotline 24078450 Fax 24082280 02014 Thermo Fisher Scientific Inc All rights reserved All trademarks are the property of Thermo Fisher Scientific and its subsidiaries unless otherwise specified TaqMan is a registered trademark of Roche Molecular Systems Inc used under permission and license Essential 8 is a trademark of Cellular Dynamics International Inc iOS is a registered trademark of Apple Inc Android is a trademark of Google Inc Sepharose is a registered trademark of GE Healthcare TRIzol is a registered trademark of Molecular Research Center Inc Facebook is a registered trademark of Facebook Inc Twitter is a registered trademark of Twitter Inc Snapchat is a registered trademark of Snapchat Inc C0011783 1214 u

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