Integrates effective genomic DNA removal with fast and sensitive reverse transcription. A simple two-step workflow generates qPCR-ready cDNA in 36 minutes.
描述
The iScript gDNA Clear cDNA Synthesis Kit integrates an effective DNase digestion step with fast and sensitive first-strand cDNA synthesis for accurate qPCR data.
This kit combines the iScript Reverse Transcription Supermix and a specially formulated DNase to effectively eliminate genomic DNA (gDNA) from purified RNA in 10 minutes. qPCR-ready cDNA free of contaminant gDNA is generated in 36 minutes.
Key Features and Benefits
- Effective genomic DNA removal — without compromising cDNA synthesis efficiency
- Proprietary DNase buffer — maintaining RNA integrity during gDNA removal
- Convenient all-in-one, single-tube supermix — for reproducible and efficient cDNA synthesis
- Ready-to-use no-RT control supermix provided — for monitoring gDNA contamination
- Superior sensitivity and reproducibility — works with a broad linear dynamic range of input total RNA (1 µg–1 pg)
- Unbiased qPCR data — consistent efficiency, sensitivity, and reproducibility in qPCR results even after gDNA removal
gDNA-free cDNA ready in 36 minutes
This kit allows you to effectively remove contaminating gDNA and reliably synthesize cDNA with minimal hands-on time in an easy-to-follow protocol, resulting in cDNA free of genomic DNA in 36 minutes.
Importance of Addressing gDNA Contamination in RT-qPCR Gene Expression
gDNA contamination in RNA samples can be amplified in RT-qPCR, biasing the results for the transcript of interest. This problem is especially acute for low expressing genes — even a few copies of gDNA contamination can shift Cq values dramatically.
Benefit of Using Reverse Transcriptase with RNase H+ in RT-qPCR Gene Expression
RNase H works to degrade RNA that is hybridized to DNA. During cDNA synthesis, the RNase H domain of the Moloney Murine Leukemia Virus (MMLV) reverse transcriptase degrades the template RNA as the cDNA is synthesized.
This activity ensures the one-to-one conversion of RNA into cDNA molecules, resulting in more accurate gene expression data. Reverse transcriptase in all iScript reverse transcription reagents exhibits RNase H activity.
Choose the Right iScript Kit for Your Needs
| Feature | Integrated gDNA Removal | Most Convenient — Just Add RNA | Maximum Input Use of up to 7.5 µg RNA | Flexible Priming Options | Value Solution |
| Product | gDNA Clear cDNA Synthesis Kit | Reverse Transcription Supermix | Advanced cDNA Synthesis Kit | Select cDNA Synthesis Kit | cDNA Synthesis Kit |
| Total Input RNA | 1 µg–1 pg | 1 µg–1 pg | 7.5 µg–100 fg | 1 µg–1 pg | 1 µg–100 fg |
| cDNA ready in | 36 min | 26 min | 21 min | 40–90 min | 26 min |
Related Products
All products listed below are compatible across real-time PCR instruments.
- SsoAdvanced™ Universal SYBR Green Supermix
Generates reliable results, due to patented Sso7d fusion protein technology, in dye-based qPCR under various challenging conditions.
- SsoAdvanced Universal Probes Supermix
Performs efficiently in challenging singleplex and duplex qPCR reactions across a broad range of reaction conditions.
- iTaq™ Universal SYBR Green Supermix
Produces consistent and reliable dye-based qPCR results.
- iTaq Universal Probes Supermix
Performs reliably and reproducibly with singleplex and duplex reactions.
More Information
Our Understanding Reverse Transcription Tutorial brings you product information, performance data, interactive tutorials, 3D animated videos, and more.
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相关资料
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Amplification Reagents and Plastics Brochure
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Reagent Comparison Guide for Real-Time PCR Brochure, Rev C
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iScript Reverse Transcription Kits Flier
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iScript™ gDNA Clear cDNA Synthesis Kit Product Information Sheet, Ver B
Gene Expression Real-Time PCR Workflow Flier
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Protocol for Multiplex RT-qPCR Analysis Using PrimePCR™ Assays, Ver A
iScript™ gDNA Clear cDNA Synthesis Kit Product Insert, Ver B