Abstract: A new method of RNA-PAP was developed that can directly amplify RNA template without additional treatment. RNA-PAP brings in a new mechanism for amplification of RNA template in which RNA-dependent DNA pyrophosphorolysis and RNA-dependent DNA polymerization are serially coupled using 3? blocked primers. Due to this serial coupling, RNA-PAP has high selectivity against mismatches on the RNA template, providing highly specific amplification of RNA template. In addition, mutant polymerases were genetically engineered for higher efficiency of RNA-dependent DNA pyrophosphorolysis and RNA-dependent DNA polymerization.

Abstract: The present invention provides a chromatography column and a method for isolating nucleic acid molecules. In one embodiment, the present invention provides a double-layer column of a first anion exchange membrane and a second serially coupled silica membrane. Upon flowing a nucleic acid-containing solution through the first anion exchange membrane, the nucleic acid binds to and then elutes from the first membrane. The eluted solution then flows serially through the second silica membrane, which the nucleic acid binds to and then elutes from. Due to this novel serial coupled double-layer principle, the present invention is particularly suitable for co-isolating RNA and DNA, for isolating nucleic acid embraced by proteins, e.g., viruses, and for isolating diluted nucleic acid in a large volume, e.g., plasma. In addition, the eluted nucleic acid is ready for downstream applications.

Abstract: A new method of RNA-PAP was developed that can directly amplify RNA template without additional treatment. RNA-PAP brings in a new mechanism for amplification of RNA template in which RNA-dependent DNA pyrophosphorolysis and RNA-dependent DNA polymerization are serially coupled using 3? blocked primers. Due to this serial coupling, RNA-PAP has high selectivity against mismatches on the RNA template, providing highly specific amplification of RNA template. In addition, mutant polymerases were genetically engineered for higher efficiency of RNA-dependent DNA pyrophosphorolysis and RNA-dependent DNA polymerization.

Abstract: A new method of RNA-PAP was developed that can directly amplify RNA template without additional treatment. RNA-PAP brings in a new mechanism for amplification of RNA template in which RNA-dependent DNA pyrophosphorolysis and RNA-dependent DNA polymerization are serially coupled using 3? blocked primers. Due to this serial coupling, RNA-PAP has high selectivity against mismatches on the RNA template, providing highly specific amplification of RNA template. In addition, mutant polymerases were genetically engineered for higher efficiency of RNA-dependent DNA pyrophosphorolysis and RNA-dependent DNA polymerization.

Abstract: The present invention provides a chromatography column and a method for isolating nucleic acid molecules. In one embodiment, the present invention provides a double-layer column of a first anion exchange membrane and a second serially coupled silica membrane. Upon flowing a nucleic acid-containing solution through the first anion exchange membrane, the nucleic acid binds to and then elutes from the first membrane. The eluted solution then flows serially through the second silica membrane, which the nucleic acid binds to and then elutes from. Due to this novel serial coupled double-layer principle, the present invention is particularly suitable for co-isolating RNA and DNA, for isolating nucleic acid embraced by proteins, e.g., viruses, and for isolating diluted nucleic acid in a large volume, e.g., plasma. In addition, the eluted nucleic acid is ready for downstream applications.