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Team:Peking S/lab/notebook/wdq
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Contents
June
| Mon | Tue | Wed | Thu | Fri | Sat | Sun |
| - | - | 1 | 2 | 3 | 4 | 5 |
| 6 | 7 | 8 | 9 | 10 | 11 | 12 |
| 13 | 14 | 15 | 16 | 17 | 18 | 19 |
| 20 | 21 | 22 | 23 | 24 | 25 | 26 |
| 27 | 28 | 29 | 30 | - | - | - |
[TOP]
6.22 - 6.25
Design primers.
6.26 - 6.27
Culture streptomycin griseus strain, which is the source of afsA, arpA, padpA.
6.28
afsA, arpA PCR using different annealing temperature, with gradient 4 ℃.
Identify them by 1% agarose gel electrophoresis.
Excise the gel slice and extract the fragments.
Second PCR using the products of gel extraction.
Electrophoresis PCR reaction system in 1.5% agarose gel.
Double digest the products of gel extraction by EcoR1-HF and Xba1.
Double digest the plasmid of B0015 by EcoR1-HF and Spe1.
Ligase afsA and arpA to the vector of B0015.
6.29
Transform the ligation reaction system.
Sythesis the genes of CAI-1 system using DNAWorks assembly and PadpA PCR.
Identify them by 1% agarose gel electrophoresis.
Excise the gel slice of pqrr, J23106+B0034+luxU, PadpA and extract the fragments.
ligase the products of gel extraction to the vector pEASY-Blunt.
6.30
Transform the ligation reaction system.
Pick six clones each plate and shave at 37℃ to amplify the bacteria.
