Biomarker Collection and Preservation for Diagnostics
Chemical Preservation of Reagents in Kits and Cartridges
PCRboost is a unique reagent that enhances end-point and reverse transcription-PCR performance by improving sensitivity and specificity during amplification of genomic DNA or RNA templates. To use, simply replace the water in your PCR with PCRboost. That's all. It is designed to enhance the amplification of DNA without the need for time consuming optimizations. PCRboost successfully amplifies thermally degraded DNA resulting in a visible amplicon that can be used for downstream applications such as cloning and sequencing.
Figure 1: Genomic DNA (gDNA) from human blood was purified using the QIAamp® DNA Blood Mini Kit., and 500ng was exposed to 70°C for 20h.
1 µl (~50ng) of thermally degraded DNA or control DNA stored at 4°C was used as template for PCR reactions. The reactions were set up using 2.5 U Taq DNA polymerase (NEB), 3μl 10x thermopol reaction buffer (NEB), 0.5µl dNTPs (10µM each nucleotide), 0.5µl each of human β-actin forward and reverse primers. The volume in the thermally degraded DNA PCR reaction tubes was brought up either in water or in PCRboost. Following PCR amplification, 10µl of each PCR reaction was run on an agarose gel. A) 500ng human genomic DNA stored at 4°C and thermally degraded at 70°C for 20h run on an agarose gel. L: 1kb ladder. B) 50ng of cold-stored (4°C) and thermally degraded DNA used as template in PCR reactions amplifying the human β-actin gene. PCR reactions were brought up to volume either in water or PCRboost. L: ladder.
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