Keith G
Biochimie. 1983 Jun;65(6):367-70. doi: 10.1016/s0300-9084(83)80159-x.
For several years most primary structure studies of ribonucleic acids have used the [32P] in vitro post-labeling techniques. We adapted our methods from the literature, and simplified them to make them accessible to any laboratory. These procedures are especially useful for preparation and purification of post labeling enzymes: T4 polynucleotide kinase, T4 RNA ligase and of gamma [32P] ATP. We developed a test tube method for 5' [32P] pCp preparation followed by tRNA labeling with T4 RNA ligase. The parameters for optimal labeling were determined. Labeling of 3.10(6) to 5.10(6) Cerenkov CPM per microgram tRNA are currently obtained.
数年来，大多数核糖核酸的一级结构研究都采用了[32P]体外后标记技术。我们借鉴了文献中的方法并加以简化，以便任何实验室都能采用。这些程序对于后标记酶（T4多核苷酸激酶、T4 RNA连接酶）以及γ[32P] ATP的制备和纯化尤为有用。我们开发了一种试管法来制备5' [32P] pCp，随后用T4 RNA连接酶对tRNA进行标记。确定了最佳标记的参数。目前每微克tRNA可获得3.10(6)至5.10(6)切伦科夫计数每分钟（CPM）的标记。
