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Team:HokkaidoU Japan/Notebook/September8
From 2010.igem.org
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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/Parts#BioBricks|1-3A]] |
* Retried September 6th transformation | * Retried September 6th transformation | ||
* pUC119 | * pUC119 | ||
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=Confirmation of pSB1C3= | =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. | 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 | + | * H buffer treated 2 uL ＋ Mighty Mix 2 uL ＋ T4 ligase 0.5 uL → Ligation → [[Team:HokkaidoU_Japan/Protocols|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 uL → Transformation | * H buffer treated 2 uL → Transformation | ||
| + | * M buffer treated 2 uL → Transformation | ||
=PCR of GFP= | =PCR of GFP= | ||
| - | * GFP reporter BBa_I13522 pSB1A2 [[Team:HokkaidoU_Japan/ | + | * GFP reporter BBa_I13522 pSB1A2 [[Team:HokkaidoU_Japan/Parts#BioBricks|2-8A]] 937 bp |
| - | * GFP protein BBa_E0840 pSB1A2 [[Team:HokkaidoU_Japan/ | + | * GFP protein BBa_E0840 pSB1A2 [[Team:HokkaidoU_Japan/Parts#BioBricks|1-12O]] 878 bp |
PCR cocktail was same as September 1st's | PCR cocktail was same as September 1st's | ||
Latest revision as of 08:14, 27 October 2010
since 13 Jul 2010
since 1 Aug 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
- M buffer treated 2 uL → Transformation
