Bio-Rad, 12004856, iScript™ Explore One-Step RT and PreAmp Kit, 50 x 50 µl rxns

50 x 50 µl reactions, contains iScript Advanced Reverse Transcriptase, 2x PreAmp Mix (dNTPs, Sso7d fusion polymerase, salts, enhancers, stabilizers, and other proprietary components), reaction booster, DNase, DNase buffer, nuclease-free water
Overview
Use the iScript Explore One-Step RT and PreAmp Kit to generate preamplified cDNA directly from RNA in a single step. This kit is suited for detecting many targets from a limited sample or for reliable detection of targets expressed at low levels. The patented Sso7d fusion protein provides unbiased target-specific preamplification of up to 100 targets. The kit also integrates an effective genomic DNA removal step, helping ensure only the transcripts of interest are detected by qPCR.
Features and Benefits
• One-step reverse transcription and preamplification — the first and only kit that eliminates protocol steps by generating preamplified cDNA from RNA in a single step
• Unbiased preamplification — can reliably profile panels of targets
• High sensitivity — enables reliable detection of genes expressed at low levels
• Preamplification control — preamplification efficacy can be easily assessed when used with PrimePCR™ long noncoding RNA (lncRNA) Arrays or custom PrimePCR Plates
• Effective genomic DNA (gDNA) removal — provides confidence that results are only for the transcript of interest
Simple Workflow
Simply create gDNA-free, preamplified cDNA by combining the reagents in this kit with RNA sample and PreAmp Assays. This kit contains all of the reagents needed to easily create an unbiased 1,000-fold amplification of cDNA.
Preamplification of Targets without Bias
Preamplification is a powerful tool to enable highly sensitive detection of a large number of targets from a limited sample. The combination of proprietary buffers and the Sso7d fusion technology enables the iScript Explore One-Step RT and PreAmp Kit to generate unbiased preamplification reactions regardless of the amount of target present.
Importance of Addressing gDNA Contamination in RT-qPCR Gene Expression
Contaminating gDNA in RNA samples can be amplified in experiments that use exonic primers, biasing the results for the transcript of interest. This problem is especially acute for low expressing genes, such as lncRNAs — even a few copies of contaminating gDNA can shift Cq values dramatically. The iScript Explore One-Step RT and PreAmp Kit addresses the issue with a specially formulated DNase that effectively eliminates gDNA from purified RNA.
Applications and Uses
• For reliable detection of targets expressed at low levels, including lncRNAs lncRNAs
• Facilitates screening a large number of genes from limited samples, especially powerful when using PrimePCR Arrays
• Ideal for single cell RT-qPCR workflows
• For highly sensitive detection when working with limited sample, including laser capture microdissections, needle biopsies, and formalin-fixed paraffin-embedded (FFPE) tissues
Packaging Options
Preamplification Product Comparison
Choose the preamplification product that best suits your needs.
• RNA is used as sample input
• Reverse transcription done using gene specific priming
• Unbiased preamplification
• Protocol minimizes hands-on time
• Compatible with PrimePCR PreAmp Control (PAQ)
• Kit integrates an effective gDNA digestion step
• Perform cDNA synthesis using any of the iScript Reverse Transcription Reagents
• cDNA is used as sample input for preamplification reaction
• Unbiased preamplification
• Flexibility with cDNA synthesis method of choice
• Archive cDNA for future studies of different genes
Related Categories
• lncRNA RT-qPCR Workflow
• Real-Time PCR Supermixes
• Real-Time PCR Detection Systems
• PrimePCR PCR Primers & Assays
• PCR Plastic Consumables
Supporting Documents
• Amplification Reagents and Plastics Brochure
• Simplified Long Noncoding RNA Discovery Brochure
• iScript Explore One-Step RT and PreAmp Kit Instruction Manual
Online Resources
• What is Real-Time PCR (qPCR)?
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