摘要
目的：建立一种简便快速的核酸浓缩回收新方法。方法：采用不同浓度的线性聚丙烯酰胺作为共沉淀载体，加入醋酸钠和无水乙醇后混匀离心，利用紫外检测和电泳方法确定对不同种类、大小和体积的核酸的回收率。结果：该方法可在１５ｍｉｎ内完成全部操作，适用于２０ｎｔ以上的单链和双链ＤＮＡ、ＲＮＡ的浓缩。对于０．１μｇ以上、大于４００ｂｐ的ＤＮＡ片段，回收率几乎达１００％。回收浓缩产物不影响后续的逆转录、ＰＣＲ、酶切和连接反应等操作。结论：所建立的方法适用于任何种类核酸的简便快速浓缩回收。
Objective: To establish a novel method for simple and rapid recovery and concentration of polynucleotides. Methods: By utilizing linear polyacrylamide at different concentrations as a carrier, sodium acetate and alcohol were added and, after mixing and centrifugation, the recovering rates of different kinds of nucleic acid were measured by UVdetector and electrophoresis. Results: Whole series of procedures could be completed within 15 min. This method is suitable to nearly any kind of polynucleotide including RNA and DNA, either single strand or double strands. A complete recovery can be achieved if the DNA fragment is longer than 400 bp and more than 0.1 μg. Samples recovered by this method can be used directly in subsequent manipulations such as reverse transcription, polymerase chain reaction, enzymatic reaction and ligation, etc. Conclusion: The method we established is suitable for simple and rapid recovery and concentration of any kind of polynucleotides.
出处
《军事医学科学院院刊》
CSCD
北大核心
1998年第1期55-57,共3页
Bulletin of the Academy of Military Medical Sciences
