PCR - SABiosciences
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1. Fluorophore Quencher Forward PCR primer Nee The fluorescence of the reporter dye is suppressed by the quencher Reverse PCR primer Amplification Assay Polymerization a_i v rr oso Ia a 2 Primer binding followed by extension e o eym Y Probe displacement and deavage Probe cleavage by Tag to free the reporter dye thus the fluorescence intensity correlates with the initial Result E sample input amounts Fluorescence P Pp a o Tag has 5 gt 3 exonuclease activity 4 r 4 PCR Products Cleavage Products Each amplicon needs a seguence specific probe cost amp time Sample to Insight Fluorescence Signal Sample to Insight Reporter Chemistries Understanding Kinetics in PCR Amplification Plot Linear scale 46 34 W 2M hb ewe wee a PM 4 349 Cycle e Plateau phase End point PCR data collection at plateau gel analysis Reactions varying due to reagent depletion amp decreased PCR efficiencies enzyme activity more product competing for primer annealing e Exponential Phase Real time PCR does early phase detection proportional to input amounts 2 dilution factor 7 evoco Characteristics of a good qPCR Assay What factors do you need to address to create a good PCR Assay Amplification efficiency 100 during exponential phase template product doubles with each cycle Sensitivity Able to detect down to reasonable quant2. I AJ 2 Annealing 60C 5 Primers 2 age Ya Sample to Insight gPCR in Action ml Polymerase 2 Annealing 60C Sample to Insight DNA Template ss or ds Polymerase MH dNTPs ara Primers 2 gPCR in Action Polymerase N k bh Polymerase art iets AO DNA Template ss or ds Polymerase eo dNTPs maa Primers 2 3 Extension 60C Sample to Insight gPCR in Action Polymerase CC COECOROAO De se DNA Template ISS Or AS Polymerase Polymerase aan dNTPs Primers 2 3 Extension 60C Pee Sample to Insight 99999 gPCR in Action QIAGEN gt 41 j Ee ee eee o A bi O A A Ak DNA Template d ss or ds Ames Polymerase E dNTPs megu Primers 2 2 a 4 Repeat 95C prum cano Sample to Insight 99999 gPCR in Action IAGEN Pi a a E e E EE E snap OOD e DNA Template O How do you make this a quantitative PCR Measure DNA amount at end of each cycle to get ratio of DNA or absolute amount if using a Bolmerace standard atap dNTPs o ara Primers 2 4 Measure amount of PCR Product E Sample to Insight 99020 Reporter chemistries QIAGEN Real Time qPCR Fluorescence Chemistry DNA binding agents n SYBR Dye Hydrolysis Probes n Dual labeled Hydrolysis Taqman probe Others such as hy
3. control gt AA Ct A Ct treated Ct control Normalized target gene expression level 2 44 Sample to Insight _ 3883s Data amp analysis Delta Delta Ct Method Amplification Plots SS GAPDH control R GAPDH treatment mea TNFa control INFo treatment GOI analysis line 0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 Cycle number AACt ACt TNFa ea GAPDH reat Act TNFO ontro GAPDH The fold change 24400 Sample to Insight Dia Data amp analysis 17 1 17 2 17 2 aPCR replicates 1 Average Ct values for all gene replicates 2 Calculate Delta Ct value GOI HKG 13 8 14 3 14 4 3 Average Delta Ct values between experiments replicates Average 14 17 4 Calculate Delta Delta Ct values Delta Ct experiment Delta Ct control DD Ct 5 00 5 Calculate Fold Change 2 Delta Delta Ct TNFa is up regulated 32 fold in the treated cells versus the control Sample to Insight Resources Support About QIAGEN Careers Ses a Hello Christine Davis Y Order TA Wishlist Cd Contact us Produets Genes amp Pathways Research Portais Technology Portais Data Analysis Center Easily interpret results The GeneGlobe Dats Analysis Center is a web resource for scientists analyzing their real time PCR or NGS dats The real time PCR modules transform threshold cycle Cr values to calculated results for gene and miRNA expression somatic muta
4. 290089 Introduction To Real Time Quantitative PCR gPCR 00000 QIAGEN 7 SA q Analysis Webinar related questions Technical Support QlAwebinarsOQIAGEN com BRCsupport giagen com Sample to Insight secas Welcome to our 4 part webinar series on qPCR 00000 QIAGEN gPCR technology overview applications dat analysis and interpretation Registration link https attendee gotowebinar com register 1255411748535841794 O Part 1 Introduction to Real Time PCR Q PCR gPCR grt PCR A Part 2 Pathway focused Gene Expression Analysis Advanced Real Time PCR Array Technology O Part 3 PCR Array Data Analysis Tutorial O Part 4 Accelerate your discovery with QIAGEN service solutions for biomarker research Sample to Insight 806888 29389 Legal Disclaimer QIAGEN a f dg nas rig ta Pa a a mma Fi j QIAGEN products shown here are intended for molecular biology applications These products are not intended for the diagnosis prevention or treatment of a disease For up to date licensing information and product specific disclaimers see the respective QIAGEN kit handbook or user manual QIAGEN kit handbooks and user manuals are available at www QIAGEN com or can be requested from QIAGEN Technical Services or your local distributor Sample to Insight 20209 How does gPCR work 009000 QIAGEN Question How far apart are the 2 cars Cars race at same sp
5. 8 m S28 338282288 oF FS PR SSE os FS 888 E nossas mr E 082393328 SP KEK KD 32288388 032454583220 ns sa 8 2 2 s2a25 83 2 qaavuaweort Positive controls Reverse transcription PCR controls Genomic DNA control Housekeeping genes J B QHDDODOHOVDDHOAVHOS RGQOHGQHHHOQOHHHAVHOHOS L Q GQHHOOHHAGQHHHOHS aDOVOVODARARDADO EII IO 7 O QQOGHOHGHHHHWHHHHH 2OQH DQHHOHOHD HHHHS OGCOCOOODOOCODOOOOQG OPOOOGOOOOOOOOOOO OOONOOOOOCOOOOOO G QHQGQHOOOHHHDDOHO 9 HQHGHHHOHHHDWBOQOHO D G9 QQSGHVDHHHDDOHOHH B QHG HHHADWDHOOOS qavawwOre nanwHEZ00a Positive PCR controls Reverse transcription controls Genomic DNA control Housekeeping genes GDC RTC PPC HKG Pre validated qPCR assays with controls Sample to Insight 29380 Want to learn more QIAGEN Visit our new Biomarker Insights Blog Just go to http biomarkerinsights giagen com Some recent topics miRNA and IncRNA serum biomarkers for lung cancer Routine blood draw biopsy of the future Liquid biopsy a necessity in the age of personalized medicine Sample to Insight 299909 Thank You for Attending 00000 QIAGEN Questions Ask now or contact Technical Support e Call 1 800 426 8157 e Email BRCsupport QIAGEN com Sample to Insight
6. CR ES Gene Expression Profiling Analysis cDNA analysis two step qRT PCR miRNA Expression Profiling Analysis TT RNA template cDNA template y v Transfer of cDNA amplifcation T qPCR products y ev y SNP Genotyping amp allelic discrimination gDNA analysis qPCR Somatic Mutation Analysis Copy Number Detection Variation Analysis Chromatin IP Quantification DNA DNA Methylation Detection re Pathogen Detection J N Amplification Viral Quantification Y y q QPCR products Sample to Insight Sample to Insight What is gPCR Applications and workflow Work Flow A Brief Look Sample RNA total mRNA Sample QC Sample QC Instrument Set up amp thermal cycling Data Output amp Analysis _ 3883s What is qPCR Applications and workflow Applications for qPCR ES Gene Expression Profiling Analysis miRNA Expression Profiling Analysis SNP Genotyping amp allelic discrimination Somatic Mutation Analysis Copy Number Detection Variation Analysis Chromatin IP Quantification DNA Methylation Detection Pathogen Detection Viral Quantification Sample to Insight 20820 gPCR for gene expression Application example QIAGEN Osteogenesis Day 16 e hMSC T4 T1 T2 T3 T4 Gene expression changes during differentiation Differentiation protocol Collect Total RNA at different time points Measure 1 HKG and 1 GOI TN
7. Fa Repeat experiment 3x biological replicates Sample to Insight 29899 gPCR for gene expression 00000 QIAGEN Work Flow Gene expression profiling y Y y SYBR y Thermocycling y Data Analysis Sample to Insight 0 22352 Factors Critical For A Successful gPCR Assay A DNA or RNA sample preparation Template quality Appropriate sample prep kits reagents Inhibitors can compromise RT or PCR B Reverse transcription to convert RNA to cDNA Choose RT kits type of RT which type of primers controls C Assay design chemistry specificity PCR efficiency amp throughput amp cost Choose validated assay or need to validate our own D Running PCR Commercial mastermix or make own primer probe master mix E Data analysis tool User friendly Streamlined data analysis module Sample to Insight 29989 RNA purity and integrity QIAGEN RNA Isolation Considerations Sample type amount Target total RNA mRNA miRNA Throughput Difficult samples stool FFPE Challenges Contamination gDNA Yield Quality Method Qiazol Column based method RNeasy Both Efficient lysis and inhibition of RNases molecular grade RNA miRNA Use a kit specific for miRNA and mRNA Sample to Insight 00060 20009 QIAGEN Sample to Insight RNA purity and integrity miRNeasy mini Kit Qiazol phen
8. Human Inflammatory Cytokines amp Receptors RT Profiler PCR Array ig 5 TI y ia pS 8 18 49 19 208 2087 1 7 0 18 10 1 A o amp y THRESHOLD CYCLE C a 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 CYCLE NUMBER Figure 4 RT Profiler PCR Arrays and RT qPCR Assays Have Sufficiently Wide Dynamic Ranges Sample to Insight 22352 Characteristics of a good gPCR Assay Sensitivity Sensitivity How many copies can my assay detect Important for low expressed genes or where there is limited sample Two Methods Method 1 Use primers to make PCR product T A clone grow up isolate guantitate and use for gPCR reactions Method 2 Use gDNA as template and use mass of gDNA to calculate copy number and assume 1 target per genome or actually calculate targets using bioinformatics 29000 Characteristics of a good gPCR Assay Specificity 00000 QIAGEN rn hb Ko N u oz ww ts z o o AGAROSE GEL Sample to Insight SINGLE DISSOCIATION CURVES BA Specificity 1 target amplified Two Methods Melt Curve analysis 1 peak 1 product Agarose gel Band at correct size 7 22352 Characteristics of a good gPCR Assay Specificity Melting Curve Analysis Plot Normalized Reporter The General Program Steps Fe ii Haa Heat to 94 to denature DNA LASG e PPK Cooling to 60 to let DNA bo Sart n E SNS
9. asor mix e qPCR Reaction Sample to Insight 00060 0009 QIAGEN Reverse Transcription Reverse Transcription Used to make cDNA copy of RNA Reagents Reverse transcriptase many different kinds dNTPs Buffers for RT Primers Random pentamers or hexamers Oligo dT Both Control RNA to monitor reverse transcription kit Important Notes RT reaction is linear Do not try to reverse transcribe too much RNA Sensitivity of GPCR step is dependent on good RT reaction Monitor RT reaction for equal efficiency across samples Sample to Insight What is in a PCR Reaction PCR Polymerase Chain Reaction Exponential Amplification of DNA in single tube All reagents in excess non limiting Components Thermostable polymerase dNTPs Primers Template Sample to Insight _ mA AAA qPCR in Action N T DNA Template ss or ds Polymerase ET dNTPs ara Primers 2 DNA Template ss or ds gPCR in Action A Polymerase Le dNTPs ara Primers 2 age Ya 1 Heat denature template 95C 2 Annealing 60C 3 Extension 60C 4 Repeat 95C Sample to Insight gPCR in Action Heat denature DNA Template ss or ds Polymerase de dNTPs 1 Heat denature template 95C ara Primers 2 age Ya Sample to Insight eegee qPCR in Action QIAGE l DNA Template Polymerase RT dNTPs
10. bridization probes o Molecular beacon and scorpion probes Sample to Insight 29089 Reporter chemistries SYBRO Green Assay 060009 QIAGEN Non fluorescent SYBR Ga a o o o SYBR I binds to double strand DNA but not s a denaturation single strand DNA Little fluorescence emitted CN O E a from SYBR I in solution o img ae SYBR upon binding to double strand DNA c o a Y gt primer annealing oo emits fluorescence very brightly o a Fluorescent SYBR The SYBR I signal intensities correlate with DNA amplified amplicon amount thus the initial sample input amounts NA TT extension BEE HERE HE ES RARE e Simple amp cost saving High Specificity Is Required when using SYBR Green since SYBR binds all double strand DNA non specific or primer dimer Sample to Insight Fluorescence Signal Sample to Insight Reporter Chemistries Understanding Kinetics in PCR Amplification Plot Linear scale 46 34 W 2M hb ewe wee a PM 4 349 Cycle e Plateau phase End point PCR data collection at plateau gel analysis Reactions varying due to reagent depletion amp decreased PCR efficiencies enzyme activity more product competing for primer annealing e Exponential Phase Real time PCR does early phase detection proportional to input amounts 2 dilution factor 22888 Reporter Chemistries Hydrolysis Based Probe Tagman Probe Assay
11. double strands anneal 3 SORA SON Slowly heat increase temp to Q Te e i 2 E AH 0 2C sec while plotting the O a PRH fluorescent signal vs E EA 50 fluorescence temperature q pe gt lt drop AN As the temp increases DNA 2 ny E na melts fluorescent signal should O 1 ON 2 AA decrease L ERCE o j aW 85 0 Significant drop in signal when Gene A Tm 77 36 Gene B Tm 78 94 See aN incite Temperature Sample to Insight 7 29352 Characteristics of a good gPCR Assay Specificity Melting Curve Analysis Plot 15 negative Derivative Reporter Single melt curve of each amplicon is required for specificity validation Derivative Reporter Rn delta F delta T the change rate Fi 65 0 70 0 75 0 80 0 85 0 90 0 25 0 Tm 78 74 Temperature C Temperature Sample to Insight 99999 Analyzing gPCR curves How to Define Baseline QIAGEN Linear Amplification Plot 08 Automated Baseline Option m A if an instrument has a adaptive baseline ng function 05 3 Manual Baseline Option a 1 Use linear view of the plot 03 2 Set up the baseline reading from 02 A cycle 2 to the cycle that 2 cycles before 01 il Ct the earliest visible amplification Baseline Iei PE 3 Usually a baseline falls in 3 15 cycles KT ST TT TT pe PD ME MAP SG Cyele Sample to Insight Analyzing gPCR curves How To Defin
12. e Threshold Log View Amplification Plot maan Use log view of amplification plot Exponentia Threshold should be higher than remy baseline higher than the noise level 1 Threshold Threshold should at LOWER 1 3 or 1 2 Oh of the linear phase of amplification Linear phase exponential phase 0 001 A Different runs across samples for the Ct same experiments should have the iii same threshold for comparison a an a SIDAD 460004046024 NADIO Cycle Sample to Insight 29920 Data Analysis Biological replicates and technical replicates QIAGEN Biological Replicates 3 different experiments Shows variability due to experiment Technical replicates 3 different measurements for same step Shows variability due to pipetting machine enzymes etc Sample to Insight E 22335 Data Analysis Housekeeping Reference Genes Any changes GOI A in control cells Ly GOI A in drug treated cells Reference gene Em Expression level remains consistent under experimental conditions different tissues E Aimed to normalize possible variations during E Sample prep handling e g use the same number of cells from a start E RNA isolation RNA quality and quantity E Reverse transcription efficiency across samples experiments E PCR reaction set up E PCR reaction amplification efficiencies Sample to Insight 22520 Data Analysis Commonly Used Housek
13. eed to finish line As car 1 crosses finish line calculate time for car 2 to finish Calculate difference in starting position mathematically d rate x time Sample to Insight How does gPCR work Question How far apart are the 2 cars Many cars how to differentiate cars of interest Sample to Insight Seminar Topics 1 What is gPCR Applications and workflow 2 QPCR for gene expression What is the change in gene expression during differentiation Factors Critical For A Successful gPCR Assay RNA purity and integrity Reverse Transcription qPCR in Action Reporter chemistries Characteristics of a good gPCR assay cr 3 PS SS Analyzing qPCR curves 10 Data amp analysis Sample to Insight What is gPCR Applications and workflow What does Real Time gPCR stand for Quantitative Polymerase Chain Reaction qPCR is a sensitive and reliable method for detection and quantification of nucleic acid DNA amp RNA levels lt is based on detection and quantification of fluorescence emitted from a reporter molecule in real time This detection occurs during the accumulation of the PCR product with each cycle of amplification Monitor the PCR reaction during early amp exponential phase where the first significant increase in the amount of PCR product correlates to the initial amount of target template Sample to Insight What is gPCR Applications and workflow Applications for qP
14. eeping Genes QIAGEN Table 7 Housekeeping genes commonly used as endogenous references Gene symbol Relative expression level Gene Human Human Mouse 18S ribosomal RNA RRN18S Actin beta ACTB Glyceraldehyde 3 phosphate dehydrogenase GAPDH Phosphoglycerate kinase 1 PGKI Peptidylprolyl isomerase A PPIA Ribosomal protein L13a RPLI3A Ribosomal protein large PO RPLPO Acidic ribosomal phosphoprotein PO Beta 2 microglobulin B2M Tyrosine 3 monooxygenase tryptophan YWHAZ 5 monooxygenase activation protein zeta polypeptide Succinate dehydrogenase complex subunit A flavoprotein Fp Transferrin receptor Aminolevulinate delta synthase 1 Glucuronidase beta Hydroxymethylbilane synthase Hypoxanthine phosphoribosyltransferase 1 TATA box binding protein Tubulin beta Tubulin beta 4 Nya indicates relative abundance of the transcripts Sample to Insight Data amp analysis 1 Average Ct values for all gene replicates 2 Calculate 4Ct value between GOI and HKG for each experiment 3 Average 4Ct values between experiments replicates 4 Calculate 4Ct values ACt experiment 4Ct control 5 Calculate Fold Change 24400 Sample to Insight 29000 Data 4 analysis 00000 QIAGEN Normalized Gene Expression Level Any changes Target Gene A in control cells Target Gene A in drug treated cells ACt Ct Target A treated Ct Ref B treated ACt Ct Target A control Ct Ref B
15. ications Liquid biopsy a new era in diagnosis Addressing challenges of NGS workflows 21 QlAsymphony Targeted NGS for cancer research Optimized NGS workflow mutation detection in cfDNA Biomedical Genomics Workbench and IVA miRNA biomarker discovery 15 miRNA overview 22 miRNA expression analysis and quantitation 29 Functional analysis of miRNA Practical hints for successful PCR 16 Novel in process monitoring qPCR tools Multiplex PCR for genotyping 23 Critical factors for multiplex qPCR Critical factors for mulitplex end point PCR 30 Streamline gene expression workflow Streamlined genotyping workflow with QlAxcel qPCR introduction 17 Coding amp non coding RNA analysis using qPCR 24 PCR array data analysis tutorial 31 Service for biomarker research 9 30 am EST 1 30pm GMT 1 00 pm EST 5 00pm GMT Register http www giagen com Knowle dge and Support Webinars 384 Well Upcoming Webinars Still searching gene by gene Learn about RT Profiler PCR Arrays Catalogued RT Profiler by pathway amp disease L OHDOHDOOHDAD DOHOOS RH BOQOHDDD DODOH HHIDOS L QQ DQHEODO OHOHADDO DGOS 1 QOQGQH OHOHAADOOSS 100008026060606068 0990850 0O10F0A00 G HD QHAQOHOHHDOHHDSOS DODVOPHPPANHHODO DOOM DADAHOO DOVOBOCODARAHHO 9 OOQHAD GOOHDADDOAGQS OVO 508 qavawwore sa sFEZO0a Asas Cr s
16. ities of template in 1 reaction 10 50 copies Specificity 1 assay 1 target no off target amplification or primer dimers e Melt curve analysis 1 peak 1 product e Agarose gel Dynamic Range Ability to detect genes with varied expression levels another judge of sensitivity e 10 to 10 copies is ideal Reproducibility Confidence in your results enables profiling of multiple genes in the same sample All lab members get the same results Technical reproducibility ensures changes seen in results are due to the biology and not the technology itself or sample handling Sample to Insight coge Characteristics of a good qPCR Assay Amplification Efficiency 00000 QIAGEN aie Ca Amplification Efficiency reliable and accurate experiment Two Methods Standard curve X axis dilution 0 1 600 1 801 1 602 1 603 1 804 1 605 1 8206 1 807 1 808 1 609 1 610 a NUMBER OF GENE COPIES Y axis Ct value THRESHOLD CycLE C Amp efficiency 10 1 slope 4 100 RN VS CYCLE Single curve analysis TW PCR Miner ta A E http miner ewindup info ver VA Gl Wl al wy son A f 1 AI di Sd DART www gene Tar i ie 18 20 22 24 26 28 50 32 38 36 38 M quantification de DART PC CYCLE NUMBER R version 1 0 xls R THRESHOLD CYCLE C Figure 4 RT Profiler PCR Arrays and RT qPCR Assays Have Sufficiently Wide Dynamic Ranges Sample to Insight Positive calls 4 100 Input RNA ng
17. ol guanidine based lysis Instant inactivation of RNases Instant end of biological activities Column cleanup Molecular biology grade RNA Archive miRNA for next project RNA purity and integrity Sample Quality Purity Quantity Spectrophotometer measure 260 280 and 260 230 m OD260 is used to calculate amount of nucleic acid oO 260 280 ratio typical minimum value 1 8 2 0 oO 260 230 ratio typical minimum value 1 7 m Low ratio may indicate a contaminant protein QIAzol Carbohydrates Glycogen m Absorbance measurements do not show integrity of RNA Integrity Denaturing RNA Agarose Gel m Usually through ribosomal bands QlAxcel Bioanalyzer m Capillary electrophoresis m Automate RNA integrity analysis m RNA integrity analysis number Agilent 2100 Bioanalyzer Sample to Insight sess Factors Critical For A Successful qPCR Assay A Templates B Primers Probes RNA Starting amount 10 1000 copies C Master Mix of NA per qPCR assay DNA polymerase For a low expressed gene need Mg 10ng equivalent of RNA per dNTPs reaction Buffer Want to start with about 100pg to Passive reference dye tug RNA Reverse Transcription D Cycling Conditions One Step or Two Step Reaction Denature gt Annealing gt Extension Denature gt Annealing Extension e 1 Tube Reaction 2 separate reactions z RT Reaction A Transfer CONA aliquers sler mix to PCR m
18. ting6h Resting6h 6 h Stimulation 6 h Stimulation Control Group Control Group Control Group Test Group 1 Test Group 1 29 08 29 02 29 27 29 89 32 02 32 13 31 96 31 15 33 83 34 22 33 09 31 57 33 95 33 26 32 65 31 3 Undetermined Undetermined Undetermined Undetermined Undetermined 29 28 84 28 53 26 67 Undetermined 37 05 Undetermined 37 28 27 33 27 11 27 31 29 54 25 52 25 6 25 81 21 26 27 12 27 21 27 12 16 77 35 53 36 21 37 66 33 49 23 28 23 16 21 34 36 32 92 35 17 36 08 34 1 33 1 25 36 25 3 25 02 96 Well Cataloged PCR Array Sample to Insight Topics Covered Today 1 What is gPCR Applications and workflow 2 QPCR for gene expression What is the change in gene expression during differentiation Factors Critical For A Successful gPCR Assay RNA purity and integrity Reverse Transcription Reporter chemistries 3 4 5 6 qPCR in Action 8 Characteristics of a good qPCR assay 9 Analyzing qPCR curves 10 Data amp analysis Sample to Insight Molecular mechanisms of neurodegeneration Host pathogen interaction Non coding RNAs amp neurodegeneration Microbiome from identification to characterization 20 Circulating biomarkers for Alzheimer s disease Innate immune system 27 Basics of sample prep in RNA research Toll like receptors in inflammation Sample to Insight cfDNA isolation and sample handling NGS introduction to technology and appl
19. tion detection and copy number measurements The NGS module supports the analysis of QIAGEN s GeneRead target enrichment panels Register on QIAGEN com to sccess this complimentary suite of tools to accelerate your data analysis and interpretation Learn more Demo gt Resources Choose format Experiment Performed Using Please choose Choose Array Choose Instrument Choose Data Analysis Type Specify CatNo Sample to Insight Investor Relations Press amp Media SiteMap Privacy Policy Trademarks amp Discialmers Cnange Language Contact QIAGEN Contact Technical Service Contact Customer Care Sunshine Act More Contacte QIAGEN 2013 15 All rights reserved Sample to Insight cones Data Analysis Tools Resources Support About QIAGEN Careers rc a Hello Christine Davis Y Order Ly Wishlist CJ Contact us v Upload data gt Analysis setup Plots amp charts gt Export data gt What s next You Selected Instructions Technology RT Profiler PCR Array Catalog Number PAHS 021Z Plate Format 96 well Want to analyze data from a different product Return to GeneGlobe Data Analysis Center Fie No file selected File must be a MS Excel Sheet in XLS format not XLSX E Does your data look like this If not please download the appropriate Excel template to format your data correctly Excel Templates for Formatting your Experimental Data E B C D E E IResting6h Res
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