NYMU Taipei/Lab Notes/2007 9 1

From 2007.igem.org

(Difference between revisions)
Line 35: Line 35:
**H2O: 14ul
**H2O: 14ul
**60°C dry bath, 1h
**60°C dry bath, 1h
 +
[[Image:NYMU_Taipei_gel_20070901_plasmid_check_1.jpg|300px]]
 +
*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

Revision as of 09:41, 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

300px

  • 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