Team:HokkaidoU Japan/Notebook/October2
From 2010.igem.org
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 |