Team:HokkaidoU Japan/Notebook/September3

From 2010.igem.org

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=[[Team:HokkaidoU_Japan/Protocols|PCR]] of [[Team:HokkaidoU_Japan/Materials_And_Methods#BioBricks|1-3A]]=
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[[Team:HokkaidoU_Japan/Protocols|Transformation]] of [[Team:HokkaidoU_Japan/Material_And_Methods#Materials_And_Methods|1-3A]] part which is originaly on pSB1C3 was succesful<br>
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[[Team:HokkaidoU_Japan/Protocols|Transformation]] of [[Team:HokkaidoU_Japan/Materials_And_Methods#BioBricks|1-3A]] part which is originaly on pSB1C3 was succesful<br>
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Thinking that linearized vector might gone bad we PCRed pSB1C3 from [[Team:HokkaidoU_Japan/Material_And_Methods#Materials_And_Methods|1-3A]]
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Thinking that linearized vector might gone bad we PCRed pSB1C3 from [[Team:HokkaidoU_Japan/Materials_And_Methods#BioBricks|1-3A]]
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Revision as of 11:32, 16 October 2010

Notebook

Resultsof yesterdays transformation

  • pSB1C3 uterly failed to produce colonies
  • pUC119 produced 20 colonies
  • colonies that should been red because of RFP insert wasn't, so there is posibility that insert wasn't there

Colony PCR on yesterdays E.coli

  • Colony PCR was done acordig to protocol
  • This day we did 20 samples

Electrophoresis

Concentration check of parts used for transformation yesterday

Electrophoresis of parts before ligation

Confirmed that DNA solution concentration of parts for Ligation was as anticipated (good?)

Lane DNA
1
2 Lambda/Hind III, EcoR I
3
4 RFP
5 pUC119
6 pSB1C3

PCR of 1-3A

Transformation of 1-3A part which is originaly on pSB1C3 was succesful
Thinking that linearized vector might gone bad we PCRed pSB1C3 from 1-3A

Reagent Amount
1-3A 1
DW 33
10x Buffer 5
2 M 4dNTPs 5
25 mM MgSO4 3
Suffix-F 1
Prefix-R 1
KOD 1
Total 50 uL
  • Extension was for120 sec
  • Purified via Microcon YM-10
  • Final volume was 43 uL
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