Chen D Y, Swerdlow H P, Harke H R, Zhang J Z, Dovichi N J
Department of Chemistry, University of Alberta, Edmonton, Canada.
J Chromatogr. 1991 Oct 18;559(1-2):237-46. doi: 10.1016/0021-9673(91)80074-q.
A low cost, 0.75-mW helium neon laser, operating in the green region at 534.5 nm, is used to excite fluorescence from tetramethylrhodamine isothiocyanate-labelled DNA fragments that have been separated by capillary gel electrophoresis. The detection limit (3 sigma) for the dye is 500 ymol [1 yoctomole (1 ymol) = 10(-24) mol] or 300 analyte molecules in capillary zone electrophoresis; the detection limit for labeled primer separated by capillary gel electrophoresis is 2 zmol [1 zeptomole (1 zmol) = 10(-21) mol]. The Richardson-Tabor peak-height encoded sequencing technique is used to prepare DNA sequencing samples. In 6% T, 5% C acrylamide, 7 M urea gels, sequencing rates of 300 bases/hour are produced at an electric field strength of 200 V/cm; unfortunately, the data are plagued by compressions. These compressions are eliminated with addition of 20% formamide to the sequencing gel; the gel runs slowly and sequencing data are generated at a rate of about 70 bases/hour.
一种低成本、功率为0.75毫瓦的氦氖激光器，在534.5纳米的绿色区域工作，用于激发经毛细管凝胶电泳分离的异硫氰酸四甲基罗丹明标记的DNA片段的荧光。该染料在毛细管区带电泳中的检测限（3σ）为500 yoctomole（1 yoctomole（1 ymol）= 10^(-24)摩尔）或300个分析物分子；经毛细管凝胶电泳分离的标记引物的检测限为2 zeptomole（1 zeptomole（1 zmol）= 10^(-21)摩尔）。采用理查森 - 塔博峰高编码测序技术制备DNA测序样品。在含6% T、5% C的丙烯酰胺、7 M尿素凝胶中，电场强度为200 V/cm时测序速率可达300个碱基/小时；不幸的是，数据存在压缩问题。在测序凝胶中加入20%甲酰胺可消除这些压缩现象；凝胶运行速度变慢，测序数据生成速率约为70个碱基/小时。
