PCR primers and library construction method for next-generation sequencing of chromaffin cell pathogenic gene

Abstract

The invention discloses a PCR primer used for building a library for chromaffin cell pathogenic gene next generation sequencing and a library building method, wherein the PCR primer is selected from nucleotides shown in SEQ ID NO:1-SEQ ID NO:70; and chromaffin cell pathogenic genes are SDHB, SDHC, SDHD, MAX, VHL, TMEM127 and RET. The library building method comprises the following steps: utilizing extracted human genome DNA as a template, adopting the primer as a specific amplification part of the primer to carry out first-round PCR amplification; diluting a product of the-first PCR amplification as a template of second-round PCR amplification; carrying out the second-round PCR amplification, namely carrying out another round of amplification by utilizing a universal primer of the first-round PCR amplification, and adding index required by a sequencing machine and a base sequence used for identifying the sequencing machine in a specific primer identification zone; and purifying a second-round sequencing product with nucleic acid, so that the second-round sequencing product can be subjected to sequencing on the sequencing machine. The library building method disclosed by the invention is designed and developed self and is extremely low in cost compared with a purchased foreign kit.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to a PCR primer and a library building method for second-generation sequencing of a chromaffin cell pathogenic gene. Background technique [0002] DNA sequencing has become an indispensable and important technology in biological research, fundamentally changing the way people study the blueprint of life. With the optimization of sequencing platform hardware and corresponding software, it is not the sequencing technology itself that hinders the progress of research, but the library construction and data analysis and interpretation related to it. [0003] 454 Life Sciences (Roche) first launched a revolutionary ultra-high-throughput genome sequencing system based on pyro-sequencing, pioneering the second-generation sequencing technology. Next-generation sequencing is also called high-throughput sequencing technology, and the basic principle is to synthesize and sequence a...

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