Membrane-mediated sample loading for automated DNA sequencing

被引:20
作者
Cassel, SM [1 ]
Guttman, A [1 ]
机构
[1] Genet BioSyst Inc, San Diego, CA 92121 USA
关键词
sample loading; DNA sequencing;
D O I
10.1002/elps.1150190823
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A new and improved sample loading method for DNA sequencing gels using oligo/polynucleotide binding membranes is described. The labeled DNA sequencing fragments were spotted onto a membrane sample loader, which was then placed in contact with the precast polyacrylamide separation gel. In this way, the time-consuming sample well loading by the tedious pipetting procedure was avoided. The spotted DNA fragments remain immobilized on the membrane until the separation process is initiated by the application of the electric field (an active DNA release mechanism). This novel technique enables sample loading outside of the separation platform, thereby allowing full utilization of various automated sample preparation and liquid handling (robotics) systems, resulting in "real" automated DNA sequencing. The loaded membranes can be stored for more than 24 hours for later use.
引用
收藏
页码:1341 / 1346
页数:6
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