Removal of Genomic DNA from RNA Preparations
Application Overview
The detection and quantification of RNA, often performed by RT-PCR and RT-qPCR, are critical in both molecular research and clinical diagnostics. However, these methods can't distinguish between cDNA and genomic DNA (gDNA), posing a risk of false positives and inaccurate RNA quantification. The removal of genomic DNA from RNA preparations is therefore essential for ensuring the reliability of these assays.
Application Overview
The detection and quantification of RNA, often performed by RT-PCR and RT-qPCR, are critical in both molecular research and clinical diagnostics. However, these methods can't distinguish between cDNA and genomic DNA (gDNA), posing a risk of false positives and inaccurate RNA quantification. The removal of genomic DNA from RNA preparations is therefore essential for ensuring the reliability of these assays.
THE PROBLEM These ENZYMEs SOLVE
Genomic DNA contamination is a pervasive issue in RT-PCR and RT-qPCR assays, often originating from the cells used in RNA extraction. While intron-spanning primers aim to amplify only cDNA, they can't fully eliminate the risk of gDNA competing for primers and probes. This competition can lead to false positives and inaccurate RNA quantification. Additionally, processed pseudogenes, which are identical to cDNA but lack introns, can also contribute to false positives.
THE PROBLEM These ENZYME SOLVES
Genomic DNA contamination is a pervasive issue in RT-PCR and RT-qPCR assays, often originating from the cells used in RNA extraction. While intron-spanning primers aim to amplify only cDNA, they can't fully eliminate the risk of gDNA competing for primers and probes. This competition can lead to false positives and inaccurate RNA quantification. Additionally, processed pseudogenes, which are identical to cDNA but lack introns, can also contribute to false positives.
THE PROBLEM These ENZYMes SOLVE
Genomic DNA contamination is a pervasive issue in RT-PCR and RT-qPCR assays, often originating from the cells used in RNA extraction. While intron-spanning primers aim to amplify only cDNA, they can't fully eliminate the risk of gDNA competing for primers and probes. This competition can lead to false positives and inaccurate RNA quantification. Additionally, processed pseudogenes, which are identical to cDNA but lack introns, can also contribute to false positives.
The Solution
Traditionally, DNase I treatment has been the go-to method for removing genomic DNA contamination. However, a faster and more efficient solution is now available: the Heat&Run gDNA removal kit. This kit offers a quick and reliable way to eliminate genomic DNA contamination, thereby enhancing the accuracy and reliability of RT-PCR and RT-qPCR assays.
By using the Heat&Run gDNA removal kit, researchers and clinicians can ensure that their RNA preparations are free from genomic DNA, leading to more accurate and reliable results.
The figure shows plots from several separate experiments. Traces of E.coli 23S DNA were found in all master mixes tested, with Cq values generally ranging from 30 – 35.
Application Background
