Team:Peking S/lab/notebook/wdq

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Contents

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June

Mon Tue Wed Thu Fri Sat Sun
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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.