Team:Tsinghua/Notebook/13 August 2010
From 2010.igem.org
Module I, DT and Fan's part:
Use the PEM② and PKC① to go on the further experiments. Double digestion.
Double digestion system:
PEM, PKC:
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).