Contaminating genomic DNA (gDNA) in RNA preparations can lead to incorrect quantification of RNA in RT-PCR or high DNA background in RNA sequencing. This is particularly an issue during the quantification of low-copy transcripts or the sequencing of small RNA samples. Removal of gDNA is often a necessity to prevent high DNA background in RNA sequencing. This protocol can be applied to RNA preparations before next-generation sequencing (NGS).
The gDNA removal kit is based on a recombinant heat-labile dsDNase (HL-dsDNase) to remove gDNA from RNA preparations. Through HL-dsDNase technology, contaminating genomic DNA is cleaved at ambient temperature and the enzyme is irreversibly inactivated upon increasing the temperature over 50° C.
Complete removal of gDNA from RNA preps
dsDNase inactivated without reducing RNA quality or quantity
Simple: Minimizes pipetting steps and reduces hands-on time
Scalable: Suitable for high-throughput experiments
Fast: Eliminate gDNA contamination in RNA samples within 20 minutes
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| Removal of gDNA in RNA preparations. (A) The gDNA removal kit removes at least 50 ng of gDNA in a 10 µL reaction volume. (B) Compared to the untreated control, no reduction in quantified cDNA (SDHA) was detected | ||
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PCR is a sensitive technique used in research and diagnostics. Taq polymerases, dNTP’s, buffer components, and primers/probes are frequently contaminated with E. coli DNA, which may lead to false positives if bacterial DNA is the target.
The PCR decontamination kit uses a dsDNase to remove contaminating DNA from mastermixes. The double-strand specific property allows decontamination in the presence of primers and probe. The dsDNase is irreversibly inactivated by heating to 60°C in the presence of DTT, ensuring that DNA template added to the mastermix after inactivation will not be digested. The mastermix can be used immediately after decontamination to run PCR reactions.
AMPIGENE® Taq polymerases have been treated to remove host DNA and additional clean-up is not necessary. The PCR decontamination kit is recommended for decontamination of primers or competitor’s PCR mixes.
“Removes DNA contamination from PCR mastermixes”
Improves target detection by reducing background
Does not affect PCR sensitivity
Optimized for PCR and probe-based qPCR mixes
Untreated and decontaminated qPCR 2x master mix was used for analysis of an E. coli gDNA 10-fold serial dilution with 5 steps. NTC samples were included, and all plots of the serial dilution show an average of three replicates. Inset: Standard curves calculated from Cq values obtained from analysis of serial dilution.
Quick and simple kit for purifying DNA from PCR, gels, and labeling reactions for a variety of downstream applications. Silica membrane technology in a mini spin column format to deliver reliable DNA purification from PCR products, agarose gels, and labeling reactions.
High performance:Recover up to 95% of DNA for fragments down to 50 bp
Comparison of DLR scores between Enzo and competitor Q’s PCR & Gel Cleanup columns
| Product | Sizes | Product Information |
| gDNA Removal Kit | 50 or 2500 Tests | Rapid removal of contaminating gDNA from RNA preps for low copy-copy transcripts or small samples |
| PCR Decontamination Kit | 100 Tests | Removal of contaminating DNA and salts in PCR mastermixes, without reduction in PCR sensitivity |
| PCR & Gel Clean-up Kit | 20, 50 or 200 Tests | For the Purification of PCR products, DNA extraction from agarose gels and removal of nucleotides, primers, enzymes, mineral oil, detergents, or dyes |
