DNA and RNA Detection Platform
DNA and RNA in biopsies usually exist with a trace amount (less than picogram/microlitter). It has been an open problem to quantify the tiny amount of DNA and RNA in biopsies, especially if the sample volumes are very limited. We therefore developed a Rolling Circule Amplification (RCA)-based DNA detection platform to solve this limitation. This platform has the capacity to sense several copies of DNA or RNA molecules in ten microlittle of reaction. The principle of this platform is that any DNA or RNA can be circulated as circular DNA. The circulized DNAs will be substrates of our prosperity Phi29 DNA polymerase and then amplified up to million folds with high fidelity. The amplified products could be further analysed with the next generation sequencing technology. The detailed process is outlined below. The sensitivity and specificity for DNA and RNA detection in our platform is better than the popular PCR technology.
Fig. 1. A Schematic of Our DNA/RNA Detection Platform. Our platform has capacity to detect any format and trace amounts of DNA and RNA from liquid biospies. The detailed was described below:
Any fragmented DNA, including FFPE.
Natural Circular DNA including:
---The known circular DNAs:
microDNA in body fluid;
Virus families with circular DNA genomes:Papillomaviridae (HPV); Polyomaviridae; Anelloviridae; Hepadnaviridae (HBV cccDNA); Circoviridae; Caulimoviridae; Geminiviridae; Nanoviridae; Adeno-associated viruses on special phase.
---The unknown or unculturable virus.
Single Cell Genomic DNA.
Any fragmented mRNA.
Natural Circular RNAs:
Hepatits Delta Virus;
cell-free DNA Library Construction.