Team:Tsinghua/Notebook/26 August 2010


Module I, group 2(b)

Run a colony PCR with universal primers. Amplify the bacteria at the same time.

PCR system (SuperMix):

 H2O	        8μl
 primer1	1μl
 primer2	1μl
 template	Some
 SuperMix	10μl
 Total	        20μl



The PEMKC 3-5 are positive. We’ll wait for the sequence result to ensure.

Module I, Group 2c

Pick several clones from the LB plate in the morning and amplify the plasmid by culturing the clone in LB.

Extract plasmid from the clone we Picked yesterday. Digest the plasmid with EcoRI and run gel to identify whether the clone is a positive one.

digestion system

plasmid            6ul
EcoRI              1ul
FD Buffer          2ul
H2O                11ul

Result: Two clones of PC are positive