Method for preparing nucleic acid library, its uses and kits

摘要

Provided are a method of constructing a nucleic acid library, a method of determining a nucleic acid sequence of a nucleic acid sample, and a kit thereof. The method of constructing the nucleic acid library includes the following steps: subjecting a nucleic acid sample to a DOP-PCR amplification, to obtain a first PCR amplification product; subjecting the first PCR amplification product to a second PCR amplification using a DOP-Amp primer, to obtain a second PCR amplification product; and subjecting the second PCR amplification product to an adaptor-ligation PCR, to obtain a third PCR amplification product, wherein the third PCR amplification product constitutes the nucleic acid library.

主权项

1. A method of constructing a nucleic acid library, comprising following steps of:
subjecting a nucleic acid sample to a first PCR amplification using a DOP primer, to obtain a first PCR amplification product; subjecting the first PCR amplification product to a second PCR amplification using a DOP-Amp primer, to obtain a second PCR amplification product; and subjecting the second PCR amplification product to an adaptor-ligation PCR using a first and second adaptor-ligation primer, to obtain a third PCR amplification product, wherein the third PCR amplification product constitutes the nucleic acid library, and wherein the DOP primer at least has a non-degenerate nucleotide region located at the 5′-end of the DOP primer and a degenerate nucleotide region downstream thereof, wherein the DOP primer has a sequence shown as SEQ ID NO: 1, wherein the DOP-Amp primer is homologous or substantially homologous to the non-degenerate oligonucleotide region located at the 5′-end of the DOP primer, wherein the 3′ end of the adaptor-ligation primers has a nucleotide sequence matching with the DOP-Amp primer; and wherein the first adaptor-ligation primer comprises P5 and the second adaptor-ligation primer comprises P7.