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06-020 Compositions and methods for amplification and cloning of near full-length viral genome samples


Xiaofeng Fan and Adrian M. Di Bisceglie


Long RT-PCR (LRP) amplification of RNA templates is sometimes difficult compared to long PCR of DNA templates. There exists a long felt need for a reliable method of replicating and amplifying sequences from long RNA templates. Polymerase chain reaction (PCR) is an indispensable technique in biomedical research. With known primer sequences, it can easily amplify a DNA target less than 3 kb but it has diminished power when the target is larger than 3 kb. This invention provides a method of producing a polydeoxyribonucleotide molecule by reverse transcriptase polymerase chain reaction wherein the polydeoxyribonucleotide molecule has a length of greater than 5,000 base-pairs is disclosed. The method involves combining two reverse transcriptases followed by two protocols of polymerase chain reaction. This method enables the amplification of large DNAs, such as viruses, from a sample while preserving genetic diversity of the large DNA. The method utilizes a proprietary reverse transcription oligonucleotide primer.

Intellectual Property Status

  • U.S. patent 7,786,294