Brumley R L, Smith L M
Department of Chemistry, University of Wisconsin, Madison 53706.
Nucleic Acids Res. 1991 Aug 11;19(15):4121-6. doi: 10.1093/nar/19.15.4121.
A horizontal polyacrylamide gel electrophoresis apparatus has been developed that decreases the time required to separate the DNA fragments produced in enzymatic sequencing reactions. The configuration of this apparatus and the use of circulating coolant directly under the glass plates result in heat exchange that is approximately nine times more efficient than passive thermal transfer methods commonly used. Bubble-free gels as thin as 25 microns can be routinely cast on this device. The application to these ultrathin gels of electric fields up to 250 volts/cm permits the rapid separation of multiple DNA sequencing reactions in parallel. When used in conjunction with 32P-based autoradiography, the DNA bands appear substantially sharper than those obtained in conventional electrophoresis. This increased sharpness permits shorter autoradiographic exposure times and longer sequence reads.
一种水平聚丙烯酰胺凝胶电泳装置已被开发出来，它能减少分离酶促测序反应中产生的DNA片段所需的时间。该装置的构造以及在玻璃板正下方使用循环冷却剂可实现热交换，其效率比常用的被动热传递方法高出约九倍。在该装置上可常规浇铸薄至25微米的无气泡凝胶。对这些超薄凝胶施加高达250伏/厘米的电场，可实现多个DNA测序反应的快速并行分离。当与基于32P的放射自显影结合使用时，DNA条带比传统电泳中获得的条带明显更清晰。这种更高的清晰度允许更短的放射自显影曝光时间和更长的序列读取长度。
