Tessier D C, Brousseau R, Vernet T
Anal Biochem. 1986 Oct;158(1):171-8. doi: 10.1016/0003-2697(86)90606-8.
Despite its unique ability to ligate single-stranded DNA molecules, T4 RNA ligase has so far seen little use in molecular biology due to long reaction times, modest yields, and apparent inability to promote ligation of long oligodeoxyribonucleotides. We describe here a set of reaction conditions which dramatically shorten the reaction time and give reproducible 40 to 60% ligation of DNA fragments of up to 40 bases in length. These improvements open promising new fields of application to T4 RNA ligase.
尽管T4 RNA连接酶具有连接单链DNA分子的独特能力,但由于反应时间长、产量不高以及明显无法促进长链寡脱氧核糖核苷酸的连接,到目前为止,它在分子生物学中的应用很少。我们在此描述了一组反应条件,这些条件可显著缩短反应时间,并使长度达40个碱基的DNA片段的连接产率达到40%至60%且具有可重复性。这些改进为T4 RNA连接酶开辟了前景广阔的新应用领域。