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Primepcr™ Assays and Panels Primepcr™ Assays and Panels We Focused on the Details So You Can Focus on What Really Matters — Your Results

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Primepcr™ Assays and Panels Primepcr™ Assays and Panels We Focused on the Details So You Can Focus on What Really Matters — Your Results Amplification: Consumables PrimePCR™ Assays and Panels PrimePCR™ Assays and Panels We focused on the details so you can focus on what really matters — your results. ■■ Expertly designed PCR primer and probe assays for ™ ™ quantitative PCR (qPCR) and Droplet Digital PCR (ddPCR ) ■■ Experimentally validated for guaranteed performance ■■ Assays for gene expression analysis, copy number variation, and mutation detection Interactive PDF. Drag cursor over text to see symbol. Table of Contents PrimePCR™ Real-Time PCR Products ...................................2–9 Why PrimePCR? ................................................................................3 Experimental Controls and Reference Gene Assays ............................4 Assay Performance Standards and Validation .....................................5 Assay Performance and Compatibility. ................................................6 PrimePCR Predesigned Panels ............................................................7 Custom Plates ....................................................................................9 Gene Expression Workflow for Real-Time PCR ................10 –13 PrimePCR™ Data Analysis ..................................................................12 Click on this icon to use the new PrimePCR Lookup Tool to find assays and panels for your genes of interest. bio-rad.com/PrimePCRLookup Droplet Digital™ PCR ..................................................................14–17 PrimePCR™ ddPCR™ Assays ..............................................................14 ddPCR™ Reagents and Software ........................................................15 Visit bio-rad.com/PrimePCR for assay and ordering information. ddPCR™ Workflow ..............................................................................16 PrimePCR™ Real-Time PCR Products Predesigned SYBR® Green Assays Transcriptome-wide human and mouse primer assays Predesigned Pathway Panels for SYBR® Green gene expression analysis are available A large selection of predesigned in 200, 1,000, or 2,500 reaction sizes. disease- and pathway-specific panels are available. DNA Templates Gene-specific synthetic DNA templates are designed to give a positive real-time PCR result when used with the corresponding gene assay. Custom PCR Plates Predesigned Probe Assays Custom-configured 96- and 384-well Transcriptome-wide probe assays for gene expression PCR plates can be ordered with ® . analysis are available in 500, 1,000, or 2,500 SYBR Green assays reaction sizes. Experimental Controls Control assays are available for reverse transcription, RNA quality, genomic DNA (gDNA) contamination, and PCR performance. PreAmp Assays Concentrated primers are available for target- specific preamplification of up to 100 SYBR® Green or probe assays. Custom Assays User-defined primer and probe sequences can be ordered. 2 PrimePCR™ Assays and Panels bio-rad.com/PrimePCR PrimePCR™ Real-Time PCR Products Why PrimePCR? Wet-Lab Validation of Every Primer Pair Wide Range of Predesigned Disease- Complete Solution for Real-Time PCR and Pathway-Specific Panels ■■ Offers guaranteed performance ■■ Expertly curated to include ■■ Aurum™ Total RNA Kits ■■ Eliminates time-consuming the most biologically relevant ■■ iScript™ cDNA Synthesis Kits gene targets optimization ■■ Supermixes for qPCR ■■ ■■ Customizable to include different Aids in minimum information ■■ PCR plates and tubes for publication of quantitative gene targets of interest ■■ Real-time PCR instruments real-time PCR experiments ■■ Integrated with CFX Manager™ (MIQE) compliance Software ■■ CFX Manager Software Assay Design ■■ Assay ■■ Avoided common ■■ Designed ■■ Avoided ■■ Maximized ■■ Compatible specificity single nucleotide intron-spanning secondary fraction of with standard confirmed by polymorphisms assays whenever structures transcript assay conditions next-generation in target regions possible in primer isoforms being sequencing annealing sites detected PrimePCR™ Assays and Panels bio-rad.com/PrimePCR 3 PrimePCR™ Real-Time PCR Products Experimental Controls and Reference Gene Assays Controls Experimental control assays and synthetic DNA templates are designed to assess the key experimental factors that may impact your real-time PCR results. 1 9 17 25 33 41 49 57 65 73 81 89 2 10 18 26 34 42 50 58 66 74 82 90/Ref 1 9 17 25 33 41 49 57 65 73 81 89 1 9 17 25 33 41 49 57 65 73 81 89 3 11 19 27 35 43 51 59 67 75 83 91/Ref 2 10 18 26 34 42 50 58 66 74 82 90/Ref DNA Contamination Control Assay 2 10 18 26 34 42 50 58 66 74 82 90/Ref 4 12 20 28 36 44 52 60 68 76 84 gDNA 3 11 19 27 35 43 51 59 67 75 83 91/Ref The PrimePCR DNA Contamination Control Assay is designed to determine if gDNA is present in a 3 1 119 1917 2725 3533 4431 5419 5597 6675 7753 831 918/Re9 f 5 13 21 29 37 45 53 61 69 77 85 PCR 4 12 20 28 36 44 52 60 68 76 84 gDNA sample at a level that may affect PCR results. 1 9 17 25 33 41 49 57 65 73 81 89 4 2 1210 2018 2826 3634 442 5520 6508 6686 7764 842 90gDN/ReAf 6 14 22 30 38 46 54 6 2 70 78 86 RQ 1 5 13 21 29 37 45 53 61 69 77 85 PCR 2 10 18 26 34 42 50 58 66 74 82 90/Ref Positive PCR Control Assay 5 3 1311 2119 2927 3735 4453 5531 6519 6697 7775 853 91PC/ReRf 7 15 23 31 39 47 55 63 71 79 87 RQ 2 The PrimePCR Positive Control Assay is designed 6 14 22 30 38 46 54 6 2 70 78 86 RQ 1 to assess how a given experimental sample may 34 112 1209 278 356 434 521 5609 678 756 834 91gDN/ReAf 6 14 22 30 38 46 54 6 2 70 78 86 RQ 1 8 16 24 32 40 48 56 64 72 80 88 RT adversely affect PCR performance. 7 15 23 31 39 47 55 63 71 79 87 RQ 2 45 123 201 298 376 445 523 601 698 776 845 gDNPCRA 7 15 23 31 39 47 55 63 71 79 87 RQ 2 8 16 24 32 40 48 56 64 72 80 88 RT RNA Quality Assay 65 143 221 3209 387 465 543 6 21 7609 787 865 RPQCR 1 8 16 24 32 40 48 56 64 72 80 88 RT The PrimePCR RNA Quality Assay is designed to determine whether RNA integrity may 6 14 22 30 38 46 54 6 2 70 78 86 RQ 1 7 15 23 31 39 47 55 63 71 79 87 RQ 2 adversely affect PCR results. Reference Gene Assays 7 15 23 31 39 47 55 63 71 79 87 RQ 2 We have suggested a set of commonly used reference8 genes16 that24 can32 be used40 individually,48 56 64 72 80 88 RT or easily screened using our preplated 96-well and 384-well reference gene panels. Reverse Transcription Control Assay 8 16 24 32 40 48 56 64 72 80 88 RT Reference gene assays may also be added to custom-designed plates. The PrimePCR Reverse Transcription Control Assay is designed to qualitatively assess the performance of the RT reaction. 4 PrimePCR™ Assays and Panels bio-rad.com/PrimePCR PrimePCR™ Real-Time PCR Products Assay Performance Standards Sensitivity Accurate detection of 20 copies Validated amplicon sequence with next-generation sequencing; minimal Specificity primer-dimer formation and gDNA cross-reactivity Amplification efficiency 90 –110% Minimum of 6 orders of magnitude; detection of a synthetic Linear dynamic range template standard curve from 20 to 20,000,000 copies R2 >0.98 All primers were validated PrimePCRറAssay Validation Report Assay Validation ™ using iScript Advanced Products used to generate validation data Real-Time PCR Instrument CFX384 Real-Time PCR Detection System PrimePCRറAssay Validation Report cDNA Synthesis Kit for Reverse Transcription Reagent iScript™ Advanced cDNA Synthesis Kit for RT-qPCR Real-Time PCR SupermixGene InformationSsoAdvanced™ SYBR® Green Supermix Partnered with Experts Experimental Sample Gene NameqPCR Human Reference Totalbreast RN cancerA 1, early onset ™ റ Gene Symbol PrimePCRBRCA1 Assay Validation Report RT-qPCR and SsoAdvanced Data Interpretation ■■ Organism Human Unique Assay ID This is a unique identifierBRCA1, that can Human be used to identify the assay in the literature and We have collaborated with Biogazelle, leaders in real-time PCR, ® Gene Summaryonline. This gene encodes a nuclear phosphoprotein that plays a role in maintaining genomic SYBR Green Supermix on Amplificationstability and Plot it also acts as a tumor suppressor. The encoded protein combines with Detected Coding Transcript(s) This is a list of the EnsemblAmplificationother transcript tumor of cDNA ID(s)suppressors thatgenerated this DNAassay from damage will 25 detect. ng sensors of universalDetails and fo signal rreference transducers RNA to form a large to design and experimentally validate gene expression assays each transcript can be foundmulti-subunit on the Ensembl protein website complex at www.ensembl.org.known as the BRCA1-associated genome surveillance complex (BASC). This gene product associates with RNA polymerase II and through ™ Amplicon Context Sequence This is the amplicon sequencethe C-terminal with additional domain base also pairs interacts added with to the histone beginnin deacetylaseg complexes. This protein an automated CFX384 Touch and/or end of the sequence.thus This plays is in a accordancerole in transcription with the DNA minimum repair information of double-stranded for breaks and the publication of real-timerecombination. quantitative PCR Mutations experiments in this (MIQE) gene are guidelines. responsible Fo rfor approximately 40% of for all human and mouse protein-coding genes details, please refer to the inheritedfollowing breastpublication, cancers "Primer and moreSequence than 80%Disclosure: of inherited A breast and ovarian cancers. Clarification of the MIQE Guidelines"Alternative (Bustinsplicing et plays al 2011). a role in modulating the subcellular localization and physiological function of this gene. Many alternatively spliced transcript variants some Real-Time PCR Detection System. Chromosome Location This is the chromosomal location of the amplicon context sequence within the of which are disease-associated mutations have been described for this gene but the genome. full-length natures of only some of these variants has been described. A related pseudogene which is also located on chromosome 17 has been identified. ■■ Assay Design Exonic: Primers sit within the same exon in the mRNA transcript and can potentially Assays provide confidence in results while eliminating time-consuming Gene Aliasesco-amplify genomic DNA.
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