高通量、半自动 Nextera 下一代测序（NGS）文库制备。

Highly Multiplexed, Semiautomated Nextera Next-Generation Sequencing (NGS) Library Preparation.

机构信息

Amyris Inc., Emeryville, CA, USA.

出版信息

Methods Mol Biol. 2020;2205:91-104. doi: 10.1007/978-1-0716-0908-8_6.

Abstract

High-throughput, inexpensive DNA sequencing is an essential component of large-scale DNA assembly operations. Using traditional and acoustic liquid-handling robotics, Illumina's Nextera Tagmentation reactions can be miniaturized and paired with custom PCR index primers to produce highly multiplexed NGS libraries for pooled sequencing. This chapter describes a high-throughput protocol that enables the simultaneous sequencing of thousands of DNA constructs in a single sequencing run at a dramatically reduced cost compared to bench-top methods.

摘要

高通量、低成本的 DNA 测序是大规模 DNA 组装操作的重要组成部分。通过使用传统和声学液体处理机器人，Illumina 的 Nextera Tagmentation 反应可以被小型化，并与定制的 PCR 索引引物配对，以产生用于 pooled 测序的高度多重化的 NGS 文库。本章描述了一种高通量方案，与台式方法相比，该方案能够在单个测序运行中同时对数千个 DNA 构建体进行测序，成本显著降低。