MULTIPLEXED LINKING PCR

摘要

The invention provides compositions and methods for rapid assembly of one or more assembled polynucleotides from a plurality of component polynucleotides, as well as methods of designing polynucleotide primer and linker sets useful in the assembly methods of the invention.

主权项

1. A method to assemble target sequences within a heterogenous population of nucleic acid molecules in a one pot reaction comprising:
a) amplifying a sample of heterogenous nucleic acid molecules containing at least two target sequences in a composition comprising at least two amplification primer pairs, to form a contiguous nucleic acid product molecule comprising each of the at least two target sequences,
wherein each of the at least two amplification primer pairs specifically amplifies one of the target sequences; and,wherein at least one amplification primer from each of the at least two amplification primer pairs comprises a linker sequence that is complementary to, and hybridizes with, a linker sequence present on one amplification primer from another one of the at least two amplification primer pairs; and, b) amplifying the contiguous nucleic acid product molecule in a composition comprising one amplification primer pair to form multiple copies of the contiguous nucleic acid product molecule.