Sartorius赛多利斯1784C----------KM

【交流】关于NEB的T4 DNA连接酶和快速连接试剂盒的常见问题及解答

液： 1X T4 DNA Ligase Buffer: [50 mM Tris-HCl, 10 mM MgCl2, 10 mM DTT, 1 mM ATP, 25 µg/ml BSA, (pH 7.5 @ 25°C)]。推荐DNA浓度(0.1 - 1 µM 的5′末端)。 使用注意：ATP是反应必须的辅因子。这一点与要求NAD作辅因子的大肠杆菌DNA连接酶不同。 如在反应体系中补加1 mM的ATP，连接反应还可以在NEB提供的四种限制性内切酶缓冲液(NEBuffers)或在T4 DNA多聚

Single Primer ("Semi-Random") P

Reamplification 40 µL H2O 10 µL 10x M-PCRB 10 µL 4 dNTPs @ 2 mM each 10 µL Pn primer @ 5 µM 10 µL Po primer @ 0.2 µM 10 µL DNA from part 1 10 µL "T/TS" @ 0.4 u Taq/µL --------- 100 µL Load glass capillaries and amplify: 30"x94°C 30

Preparation of Carboxypeptidase Y and Its Properties

is immersed in a water bath at 30℃ during kneading of the mixture. After liquefaction, add 600ml of distilled water, the mixture is stirred. The pH of the mixture is adjusted to 7.0 with 1 M NaOH under vigorous stirring, and the whole is kept at 25