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Team:HokkaidoU Japan/Notebook/October2
From 2010.igem.org
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* Results showed no insertion | * Results showed no insertion | ||
** but when result came, had already done miniprep and prepared Sequencing Master Mix | ** but when result came, had already done miniprep and prepared Sequencing Master Mix | ||
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== Miniprep == | == Miniprep == | ||
* Used Qiagen kit | * Used Qiagen kit | ||
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|style="border-top:1px solid #996;"|'''121.25''' | |style="border-top:1px solid #996;"|'''121.25''' | ||
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| + | [[Image:HokkaidoU Japan 20101002c.JPG|200px|right|thumb]] | ||
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Revision as of 15:10, 2 October 2010
since 13 Jul 2010
since 1 Aug 2010
Colony PCR
- Performed Colonie PCR for 3 colonies which were incubated over night
- Colony numbers were: 1, 2 and 3
- Results showed no insertion
- but when result came, had already done miniprep and prepared Sequencing Master Mix
Miniprep
- Used Qiagen kit
- Melted in 50 uL TE instead of H2O
Preparation for Sequencing
- Mixed as shown in the table below
5x Sequencing Buffer 1.5uL 24.75 uL Ready Reaction Premix 1 uL 16.5 H2O 5 uL 80 uL total 7.5/sample
| Reagent | Amount | Amount for 16.5 |
|---|---|---|
| 5x Sequencing Buffer | 1.5uL | 24.75 uL |
| Ready Reaction Premix | 1 uL | 16.5 |
| H2O | 5 uL | 80 uL |
| Total | 7.5/sample | 121.25 |
