Sartorius赛多利斯16533--------GUKM

【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

a biased ratio of Po and Pn. Make 3 reactions with varying [Mg++]: 15 µL H2O 3 µL 10x H, M or L-PCRB 3 µL 4 dNTPs @ 2 mM each 3 µL Po @ 5 µM 3 µL template diluted to about 1 ng/µL 3 µL "T/TS" @ 0.4 u Taq/µL -------- 30 µL

Screening lambda cDNA or genomic libraries

need 6.6 ml top agarose (use a bigger tube and a 50-55° water bath).to titer (on 100 mm plates):Put various dilutions of phage in 100 µl 1X phage buffer.Always do a no phage control! This will help you distinguish between dilutions that give you no phage (look