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文献和实验of a master mix containing water, buffer, dNTPs, primers and Taq DNA Polymerase in a single tube, which can then be aliquoted into individual tubes. MgCl2 and template DNA solutions are then added. This method of setting reactions minimizes the possibility
Amplification and Labelling with Cy dyes
mix 1 (for 2 reactions): 2ml 5X T7 Sequenase buffer 3ml 10mM dNTP mix 1.5ml 0.1M DTT 3ml 500mg/ml BSA 0.6ml 13U/ml T7 Sequenase Reaction mix 2 (for 2 reactions): 1.4ml Sequenase dilution buffer 0.6ml 13U/ml T7 Sequenase Program themalcycler
5. 不受粘末端或平末端的限制6. 超简洁反应体系配制:只需将克隆片段与线性化的载体按一定比例加入Lightening Assembly Master Mix即可7. 超快速:克隆连接反应只需10-15分钟较传统克隆方法,操作步骤更少,所需试剂更少,所需时间大大缩短。8. PCR产物可直接与表达载体连接,无需酶切或连入克隆载体。应用范围:1. 平末端和粘末端DNA片段克隆。 2. 1到多个DNA片段克隆。3. 点突变。 4. 长片段克隆。5. DNA片段上有和载体相同的酶切
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