Team:HokkaidoU Japan/Notebook/September8
From 2010.igem.org
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- | =Colony PCR= | + | =[[Team:HokkaidoU_Japan/Protocols|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 | 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) | ||
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* H buffer treated 2 uL + Mighty Mix 2 uL + T4 ligase 0.5 uL → Ligation → Transformation | * 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 | * M buffer treated 2 uL + Mighty Mix 2 uL + T4 ligase 0.5 uL → Ligation → Transformation | ||
- | * H buffer treated 2 | + | * H buffer treated 2 uL → [[Team:HokkaidoU_Japan/Protocols|Transformation]] |
* H buffer treated 2 uL → Transformation | * H buffer treated 2 uL → Transformation | ||
Revision as of 09:18, 2 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 tion 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