Slatko Barton E, Kieleczawa Jan, Ju Jingyue, Gardner Andrew F, Hendrickson Cynthia L, Ausubel Frederick M
New England Biolabs, Inc., Ipswich, Massachusetts, USA.
Curr Protoc Mol Biol. 2011 Oct;Chapter 7:Unit7.2. doi: 10.1002/0471142727.mb0702s96.
Beginning in the 1980s, automation of DNA sequencing has greatly increased throughput, reduced costs, and enabled large projects to be completed more easily. The development of automation technology paralleled the development of other aspects of DNA sequencing: better enzymes and chemistry, separation and imaging technology, sequencing protocols, robotics, and computational advancements (including base-calling algorithms with quality scores, database developments, and sequence analysis programs). Despite the emergence of high-throughput sequencing platforms, automated Sanger sequencing technology remains useful for many applications. This unit provides background and a description of the "First-Generation" automated DNA sequencing technology. It also includes protocols for using the current Applied Biosystems (ABI) automated DNA sequencing machines.
从20世纪80年代开始,DNA测序自动化极大地提高了通量,降低了成本,并使大型项目能够更轻松地完成。自动化技术的发展与DNA测序其他方面的发展并行:更好的酶和化学方法、分离和成像技术、测序方案、机器人技术以及计算方面的进步(包括带有质量分数的碱基识别算法、数据库开发和序列分析程序)。尽管出现了高通量测序平台,但自动化桑格测序技术在许多应用中仍然有用。本单元提供了“第一代”自动化DNA测序技术的背景和描述。它还包括使用当前应用生物系统公司(ABI)自动化DNA测序仪的方案。