聚丙烯酰胺电泳条带中微量DNA片段的回收与再扩增方法的改进

Improved methods to recover and reamplify trace DNA fragments from the polymorphic PAGE band

摘要:
目的研究聚丙烯酰胺凝胶电泳(PAGE)条带中微量DNA片段的回收和再扩增.
方法比较了4种方法溶解聚丙烯酰胺凝胶的效果和回收微量核酸的效率.
结果和结论 Bioteke溶胶/结合液能获得极高的DNA片段回收率，可从仅含1.0~7.5 ng核酸的PAGE胶中有效回收到足量的片段；再扩增的效果与回收效率呈正相关；将液态混合液与杂质分离是成功进行再扩增的关键.本研究构建的一套快速、高效的PAGE多态性条带回收及再扩增的方法为分析生物遗传多样性成因提供了新途径.

Abstract:
Objective To optimize the methods of recovery and reamplification of trace DNA fragments from polymorphic polyacrylamide gel electrophoresis (PAGE) bands.
Method Effects of four methods of dissolving polyacrylamide gel and recovery efficiencies of trace nucleic acids were obtained.
Result and conclusion Dissolving/Bing buffer from Bioteke kit showed high recovery rates of DNA fragments from PAGE gels with only 1.0-7.5 ng nucleotide. Effects of amplification positively correlated with recovery efficiencies. The separation of liquid mixture and impurities was the key to successful amplification. This research forms a fast, efficient DNA recycling and amplification method from polymorphism PAGE, which provides a new approach to analyze the causes of genetic diversity.