Nishikawa T, Kambara H
Central Research Laboratory, Hitachi Ltd., Tokyo, Japan.
Electrophoresis. 1992 Aug;13(8):495-9. doi: 10.1002/elps.11501301104.
A high-resolution separation of DNA bands is achieved by electrophoresis with a long gel in DNA base sequencing using fluorescence detection. We separate 760 and 761 base DNA fragments using the 93 cm migration electrophoresis optimized for the separation of DNA bands. A T7 DNA polymerase and an Mn++ buffer are used in sequencing reactions to obtain fluorescence peaks of uniform strength, and the peak areas in the spectrum are used for recognizing the peak number in a cluster of successive peaks. This method is successfully applied to the DNA fragment spectrum obtained by 93 cm migration electrophoresis, which results in a single-band differentiation of bands of 1040 base DNA.
在使用荧光检测的DNA碱基测序中，通过在长凝胶上进行电泳可实现DNA条带的高分辨率分离。我们使用针对DNA条带分离优化的93厘米迁移电泳来分离760和761个碱基的DNA片段。在测序反应中使用T7 DNA聚合酶和Mn++缓冲液以获得强度均匀的荧光峰，并利用光谱中的峰面积来识别连续峰簇中的峰数。该方法成功应用于通过93厘米迁移电泳获得的DNA片段光谱，实现了1040个碱基DNA条带的单带区分。
