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|
All products listed below are compatible across real-time PCR instruments.
Generates reliable results, due to patented Sso7d fusion protein technology, in dye-based qPCR under various challenging conditions.
Performs efficiently in challenging singleplex and duplex qPCR reactions across a broad range of reaction conditions.
Produces consistent and reliable dye-based qPCR results.
Performs reliably and reproducibly with singleplex and duplex reactions.
Our Understanding Reverse Transcription Tutorial brings you product information, performance data, interactive tutorials, 3D animated videos, and more.
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