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Team:HokkaidoU Japan/Notebook/September8
From 2010.igem.org
(Difference between revisions)
(→コロニーPCR) |
(→pSB1C3の確認) |
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| Line 8: | Line 8: | ||
* pUC119 | * pUC119 | ||
| - | = | + | =Confirmation of pSB1C3= |
| - | 1-3A（RFP | + | 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 | + | * H buffer treated 2 uL ＋ Mighty Mix 2 uL ＋ T4 ligase 0.5 uL → Ligation → Transformation |
| - | * M | + | * M buffer treated 2 uL ＋ Mighty Mix 2 uL ＋ T4 ligase 0.5 uL → Ligation → Transformation |
| - | * H | + | * H buffer treated 2 uL → Transformation |
| - | * H | + | * H buffer treated 2 uL → Transformation |
=GFPのPCR= | =GFPのPCR= | ||
Revision as of 18:00, 27 September 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 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
GFPのPCR
- GFP reporter BBa_I13522 pSB1A2 2-8A 937bp
- GFP protein BBa_E0840 pSB1A2 1-12O 878bp
