The free solution mobility of DNA
The free solution mobility of DNA
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DOI：
10.1002/(sici)1097-0282(199711)42:6
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发表时间：
1997-11-01
期刊：
影响因子：
2.9
通讯作者：
Righetti, PG
中科院分区：
文献类型：
--
作者：
Stellwagen, NC;Gelfi, C;Righetti, PG
DNP的自由溶液迁移率已通过毛细管电泳在最常用于DNA凝胶电泳的两种缓冲液中测量; Tris-硼酸盐-EDTA（TEE）和Tris-乙酸盐-EDTA（TAE）。用两种新型丙烯酰胺单体N-丙烯酰氨基乙氧基乙醇或N-丙烯酰氨基丙醇中的任一种的聚合物涂覆毛细管，这两种单体在碱性pH下都是稳定的，并且有效地消除了由于毛细管壁引起的电内渗、贵族化。在25 ℃时，DNA在TAE缓冲液中的自由溶液迁移率为（3.75 +/- 0.04）× 10（-4）cm（2）V-1 s（-1）。与DNA浓度、样品大小、电场强度和毛细管涂层无关，并且与文献中的其他值良好一致。自由溶液迁移率与DNA分子量无关，从类似的400个碱基对到48.5千碱基对，但随着较小片段分子量的降低而单调降低。令人惊讶的是，DNA在TEE缓冲液中的自由溶液迁移率为（4.5 +/- 0.1）x 10（-4）cm（2）V-1 s（-1），比在TAE缓冲液中观察到的高约20%，可能是因为非特异性硼酸盐-脱氧核糖复合物的形成。(C)John Wiley & Sons，Inc.
The free solution mobility of DNP, has been measured by capillary electrophoresis in the two buffers most commonly used for DNA gel electrophoresis,;Tris-borate-EDTA (TEE) and Tris-acetate-EDTA (TAE). The capillaries were coated with polymers of either of two novel acrylamide monomers, N-acryloylaminoethoxethanol or N-acryloylaminopropanol, both of which are stable at basic pH and effectively eliminate the electroendosmotic,nobility due to the capillary walls. The free solution mobility of DNA in TAE buffer was found to be (3.75 +/- 0.04) x 10(-4) cm(2) V-1 s(-1) at 25 degrees C. independent of DNA concentration, sample size, electric field strength, and capillary coating, and in good agreement with other values in the literature. The free solution mobility was independent of DNA molecular weight from similar to 400 base pairs to 48.5 kilobase pairs, but decreased monotonically with decreasing molecular weight for smaller fragments. Surprisingly, the free solution mobility of DNA in TEE buffer was found to be (4.5 +/- 0.1) x 10(-4) cm(2) V-1 s(-1), about 20% larger than observed in TAE buffer, presumably because of the formation of nonspecific borate-deoxyribose complexes. (C) 1997 John Wiley & Sons, Inc.