NYMU Taipei/Lab Notes/2007 8 30

< NYMU Taipei/Lab Notes(Difference between revisions)

Latest revision as of 07:27, 1 September 2007

Objective: RBS + cinR
Objective: RBS + HSL
Enzyme Digestion (RBS)
- RBS: 11ul
- SpeI: 1ul (10,000 U/ml)
- PstI: 1ul (20,000 U/ml)
- 10x buffer: 2, 2ul
- 10x BSA: 2ul
- H2O: 3ul
- Total: 20ul
- 37°C, 2h
- Heat inactivation: 65°C 20min
Enzyme Digestion (cinR)
- cinR: 11ul
- XbaI: 1ul (20,000 U/ml)
- PstI: 1ul
- 10x buffer: 3, 2ul
- 10x BSA: 2ul
- H2O: 3ul
- Total: 20ul
- 37°C, 2h
- Heat inactivation: 65°C 20min
Enzyme Digestion (HSL)
- HSL: 4ul
- XbaI: 1ul
- PstI: 1ul
- 10x buffer: 3, 2ul
- 10x BSA: 2ul
- H2O: 10ul
- Total: 20ul
- 37°C, 2h
- Heat inactivation: 65°C 20min
Plasmid extraction: H_IGFBP3, H_IGFBP7, H_INS, M_IDE
- From 5ml LB medium to 50ul EB
Ligation (RBS+cinR)
- RBS: 1ul
- cinR: 1ul
- 10x buffer: 1ul
- T4 ligase (Promega): 1ul
- H2O: 6ul
Ligation (RBS+HSL)
- RBS: 1ul
- HSL: 1ul
- 10x buffer: 1ul
- T4 ligase (Promega): 1ul (3 U/ul)
- H2O: 6ul
Transformation
- Competent cell: 50ul
- DNA: 2ul

@@ Line 4: / Line 4: @@
 *Enzyme Digestion (RBS)
 **RBS: 11ul
-**SpeI: 1ul
+**SpeI: 1ul (10,000 U/ml)
-**PstI: 1ul
+**PstI: 1ul (20,000 U/ml)
 **10x buffer: 2, 2ul
 **10x BSA: 2ul
@@ Line 14: / Line 14: @@
 *Enzyme Digestion (cinR)
 **cinR: 11ul
-**XbaI: 1ul
+**XbaI: 1ul (20,000 U/ml)
 **PstI: 1ul
 **10x buffer: 3, 2ul
@@ Line 44: / Line 44: @@
 **HSL: 1ul
 **10x buffer: 1ul
-**T4 ligase (Promega): 1ul
+**T4 ligase (Promega): 1ul (3 U/ul)
 **H2O: 6ul
+*Transformation
+**Competent cell: 50ul
+**DNA: 2ul
 ----
-by 立翔
+==Who attend?==
+* 立翔