Team:Tsinghua/Notebook/13 August 2010


Module I, group 2(b)

Use the PEM② and PKC① to go on the further experiments. Double digestion.

Double digestion system:


 H2O	         25.5μl
 buffer tango	 10μl
 plasmid	 10μl
 EcoRI	         1.5μl
 BamHI	         3μl
 Total	         50μl

Run a gel and cut to get the KC fragments and PEM plasmid. Ligate at 22℃ for 20min.

Ligation system:

 H2O	        11.8μl
 10×buffer	2μl
 KC	        3μl
 PEM	        2.2μl
 T4 ligase	1μl
 Total	        20μl

Transform the ligation product into bacterial cells immediately. Spread about 100μl of the resulting solutions on LB plates (with 0.1% ampicillin).