Team:Tsinghua/Notebook/26 August 2010
From 2010.igem.org
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
result
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