Wilson R K, Yuen A S, Clark S M, Spence C, Arakelian P, Hood L E
Division of Biology, California Institute of Technology, Pasadena 91125.
Biotechniques. 1988 Sep;6(8):776-7, 781-7.
We report here a system for automation of the dideoxynucleotide DNA sequencing method. The system consists of a Beckman Biomek 1000 robotic workstation which has been modified by the addition of a thin heater/cooler block directly on the instrument table. The heater/cooler block, which is regulated by a user-specified program integrated into the Biomek software, facilitates the use of both single- and double-stranded DNA sequencing procedures. Using this system, we are able to perform 24 sets of dideoxynucleotide DNA sequencing reactions in approximately 45 min. The reactions are performed in 96-well microtiter plates, using a prepared reagent pack which includes primer, enzyme, nucleotide mixes and radioactive label. Using standard gel electrophoresis technology, we are able to resolve over 500 nucleotides per sequencing reaction in about 5 h. Currently, we are able to perform three runs of 24 samples each, with subsequent gel analysis per 8-h period. Excluding autoradiography, this represents a daily data output of 36,000 base pairs.
我们在此报告一种用于双脱氧核苷酸DNA测序方法自动化的系统。该系统由一台贝克曼Biomek 1000机器人工作站组成，通过在仪器台上直接添加一个薄的加热/冷却块进行了改装。加热/冷却块由集成到Biomek软件中的用户指定程序调节，便于单链和双链DNA测序程序的使用。使用该系统，我们能够在约45分钟内进行24组双脱氧核苷酸DNA测序反应。反应在96孔微量滴定板中进行，使用包含引物、酶、核苷酸混合物和放射性标记的预制试剂包。使用标准凝胶电泳技术，我们能够在约5小时内每个测序反应解析超过500个核苷酸。目前，我们能够每8小时进行三次每组24个样本的运行，并随后进行凝胶分析。不包括放射自显影，这代表每天36000个碱基对的数据输出。
