Hebenbrock K, Williams P M, Karger B L
Barnett Institute, Northeastern University, Boston, MA 02115, USA.
Electrophoresis. 1995 Aug;16(8):1429-36. doi: 10.1002/elps.11501601236.
Capillary electrophoresis (CE), using a replaceable polymer matrix and two-dye laser-induced fluorescence has been applied to single strand conformational polymorphism (SSCP). Two-dye laser-induced fluorescence has been used for improved strand identification over a single-dye approach. Conditions suitable in the capillary format for rapid separation and high resolution have been explored. The influence of separation parameters such as temperature and matrix composition on separation in SSCP was first determined. Short analysis times allowed for fast screening of optimal separation conditions of the sample. Based on these results, the two strands of a standard 255 bp fragment of the lacI gene were resolved within 25 min with replaceable linear polyacrylamide as a separation matrix. The method was then applied to the detection of different mutations, in the presence of wild type, of a 276 bp fragment of the insulin-like growth factor 1-binding protein 1 (IGF1-BP3) gene.
毛细管电泳（CE）结合可替换的聚合物基质和双染料激光诱导荧光技术已应用于单链构象多态性（SSCP）分析。与单染料方法相比，双染料激光诱导荧光技术用于改善链的识别。已探索了适用于毛细管形式的快速分离和高分辨率的条件。首先确定了诸如温度和基质组成等分离参数对SSCP分离的影响。短分析时间允许快速筛选样品的最佳分离条件。基于这些结果，以可替换的线性聚丙烯酰胺作为分离基质，在25分钟内解析了lacI基因标准255 bp片段的两条链。然后将该方法应用于检测胰岛素样生长因子1结合蛋白1（IGF1-BP3）基因276 bp片段在野生型存在下的不同突变。
