Team:Tsinghua/Notebook/9 October 2010

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(Difference between revisions)
(Module I, Group 2c)
(Module I, Group 1A)
 
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== Module I, Group 1A ==
== Module I, Group 1A ==
Colony pcr to screen the clones in which landing pad has successfully integrated into genome, using up primers from the plasmid and down primers from the genome.
Colony pcr to screen the clones in which landing pad has successfully integrated into genome, using up primers from the plasmid and down primers from the genome.
 +
Select the positive clones to amplify in LB media with 0.03% tetracyclin.
 +
== Module I, Group 2c ==
== Module I, Group 2c ==
After purification, ligation reaction took place in a mixed system as following, respectively.
After purification, ligation reaction took place in a mixed system as following, respectively.

Latest revision as of 15:35, 27 October 2010

Module I, Group 1A

Colony pcr to screen the clones in which landing pad has successfully integrated into genome, using up primers from the plasmid and down primers from the genome. Select the positive clones to amplify in LB media with 0.03% tetracyclin.

Module I, Group 2c

After purification, ligation reaction took place in a mixed system as following, respectively.

eGFP                 3ul
PSB1C13              2ul
T4 ligase(NEB)       1ul
T4 ligation Buffer   2ul
H2O                  12ul

mCherry              2ul
PSB1C13              2ul
T4 ligase(NEB)       1ul
T4 ligation Buffer   2ul
H2O                  13ul

Kan                  3ul
PSB1C13              2ul
T4 ligase(NEB)       1ul
T4 ligation Buffer   2ul
H2O                  12ul

Incubate at 16 centidegree overnight.