NYMU Taipei/Lab Notes/2007 9 1
From 2007.igem.org
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*Well 6 7: "RBS+cinR 1" SpeI, PstI | *Well 6 7: "RBS+cinR 1" SpeI, PstI | ||
*Well 8: "RBS+cinR 1" BsaBI | *Well 8: "RBS+cinR 1" BsaBI | ||
| + | *Comment | ||
| + | **There seems to be some incomplete digestion of RBS+HSL 2, possibly result from too many DNA. | ||
| + | ** | ||
Revision as of 11:12, 1 September 2007
- Plasmid extraction: RBS+cinR 1, RBS+HSL 1, RBS+HSL 2, RBS+HSL 3 from 5ml LB culture to 50ul EB
- The concentration was measured with spectrophotometer at 260nm
- RBS+cinR 1: 8ug/ul
- RBS+HSL 1: 3ug/ul
- RBS+HSL 2: 8ug/ul
- RBS+HSL 3: 6ug/ul
- Enzyme digest RBS+cinR 1
- RBS+cinR 1: 14ul
- SpeI: 1ul
- PstI 1ul
- 10x buffer 2: 2ul
- 10x BSA: 2ul
- 37°C water bath, 2h
- Heat inactivation: 65°C, 20min
- Enzyme digest RBS+HSL 2
- RBS+HSL 2: 14ul
- XbaI: 1ul
- PstI: 1ul
- 10x buffer 3: 2ul
- 10x BSA: 2ul
- 37°C water bath, 2h
- Heat inactivation: 65°C, 20min
- Enzyme digest RBS+HSL 3
- RBS+HSL 3: 14ul
- XbaI: 1ul
- PstI: 1ul
- 10x buffer 3: 2ul
- 10x BSA: 2ul
- 37°C water bath, 2h
- Heat inactivation: 65°C, 20min
- Check RBS+cinR 1 with BsaBI digestion
- RBS+cinR 1: 3ul
- BsaBI: 1ul
- 10x buffer 2: 2ul
- H2O: 14ul
- 60°C dry bath, 1h
- Well 1: 100pb ladder
- Well 2 3: "RBS+HSL 2" XbaI, PstI
- Well 4 5: "RBS+HSL 3" XbaI, PstI
- Well 6 7: "RBS+cinR 1" SpeI, PstI
- Well 8: "RBS+cinR 1" BsaBI
- Comment
- There seems to be some incomplete digestion of RBS+HSL 2, possibly result from too many DNA.
