Continuous, on-line DNA sequencing using oligodeoxynucleotide primers with multiple fluorophores.
Continuous, on-line DNA sequencing using oligodeoxynucleotide primers with multiple fluorophores.
复制标题
使用具有多个荧光团的寡脱氧核苷酸引物进行连续在线 DNA 测序。
DOI：
--
复制
发表时间：
1988
影响因子：
11.1
通讯作者：
J. Ruth
中科院分区：
文献类型：
--
作者：
J. Brumbaugh;L. Middendorf;D. Grone;J. Ruth
描述了一种通过使用双荧光素化引物和激光激发进行DNA测序的方法。已经确定了双脱氧方案，其提供了从碱基1开始的600个碱基的序列，在单次加载中误差小于1%。电泳在20 W下进行，用扫描荧光检测器在距离上样孔底部24 cm处检测条带。波段在电视屏幕上以二维成像。通过使用简单的软件程序，可以手动或半自动地从电视屏幕上读取序列。该系统允许读取更多的碱基，错误率低于任何其他已报道的自动测序方法。
A method for sequencing DNA by using a difluoresceinated primer and laser excitation is described. Dideoxy protocols have been determined that provide sequences for 600 bases starting with base 1 with less than 1% error in a single load. Electrophoresis is at 20 W and the bands are detected 24 cm from the bottom of the loading well with a scanning fluorescence detector. Bands are imaged on a TV screen in two dimensions. The sequences can be read from the TV screen manually or semiautomatically by using a simple software program. The system allows more bases to be read with a lower error rate than any other reported automated sequencing method.