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From 2011.igem.org

Contents 1 summary 2 June 2.1 6.22 - 6.25 2.2 6.26 - 6.27 2.3 6.28 2.4 6.29 2.5 6.30

summary

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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.