Team:HokkaidoU Japan/Notebook/September8
From 2010.igem.org
(Difference between revisions)
(→Confirmation of pSB1C3) |
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Checked if primers PS-R, EX-F were good to use for colony PCR and also checked if insert was realy in plasmids | Checked if primers PS-R, EX-F were good to use for colony PCR and also checked if insert was realy in plasmids | ||
* September 6th transformation (digestion was done with H buffer×2 and M buffer×2, 2 version) | * September 6th transformation (digestion was done with H buffer×2 and M buffer×2, 2 version) | ||
- | * [[Team:HokkaidoU_Japan/ | + | * [[Team:HokkaidoU_Japan/Materials_And_Methods#BioBricks|1-3A]] |
* Retried September 6th transformation | * Retried September 6th transformation | ||
* pUC119 | * pUC119 | ||
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=PCR of GFP= | =PCR of GFP= | ||
- | * GFP reporter BBa_I13522 pSB1A2 [[Team:HokkaidoU_Japan/ | + | * GFP reporter BBa_I13522 pSB1A2 [[Team:HokkaidoU_Japan/Materials_And_Methods#BioBricks|2-8A]] 937bp |
- | * GFP protein BBa_E0840 pSB1A2 [[Team:HokkaidoU_Japan/ | + | * GFP protein BBa_E0840 pSB1A2 [[Team:HokkaidoU_Japan/Materials_And_Methods#BioBricks|1-12O]] 878bp |
PCR cocktail was same as September 1st's | PCR cocktail was same as September 1st's |
Revision as of 11:33, 16 October 2010
Colony PCR
Checked if primers PS-R, EX-F were good to use for colony PCR and also checked if insert was realy in plasmids
- September 6th transformation (digestion was done with H buffer×2 and M buffer×2, 2 version)
- 1-3A
- Retried September 6th transformation
- pUC119
Confirmation of pSB1C3
Because pSB1C3 PCRed from 1-3A(RFP reporter) was used there was posibility of contamination by template which would also produce red colonies.
- H buffer treated 2 uL + Mighty Mix 2 uL + T4 ligase 0.5 uL → Ligation → Transformation
- M buffer treated 2 uL + Mighty Mix 2 uL + T4 ligase 0.5 uL → Ligation → Transformation
- H buffer treated 2 uL → Transformation
- H buffer treated 2 uL → Transformation