Automated Sanger dideoxy sequencing reaction protocol

FEBS Letters

Schwager, C.¹  Zimmermann, J.¹  Stegemann, J.¹  Voss, H.¹  Ansorge, W.¹ 

[1] European Molecular Biology Laboratory, Postfach 10. 2209 Heidelberg, FRG

关键词: DNA sequencing;    Dideoxy reaction;    Automated sequencing;    Fluorescent label;    Radioactive label;   

DOI  :  10.1016/0014-5793(88)80477-0

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The protocol for Sanger dideoxy chain termination reactions in DNA sequencing is tedious and prone to errors due to the repetitive character of the pipetting steps. An industrial robot, with the addition of a few simple parts, was programmed to automate the dideoxy sequencing reactions. The system is set up in a short time for routine operation and it is faster and more reliable than a human operator. It is flexible and allows variations and optimization of the standard procedure. Disposable microtiter plates at a controlled temperature are used. In one reaction cycle (about 50 min) up to 48 templates are processed. Up to 450 bases were resolved in automated DNA sequencing on samples prepared by the robot. The protocol is applicable to fluorescent as well as to radioactive labeling.

【 授权许可】

Unknown