IGEM:Peking/2007/Count-Conjugation-Notebook/2007-8-16
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Tandem OriT by Qu Mingzhi
Amplification Culture of Ori-T
- select Positive OriT-pEASY-3 Colonies from Plate,Culture in liquid LB,waiting for mini-prep.
mini-prep E0240
- using Transgen mini plasmid puriflication kit.
- 50µL after purflication.
mini-prep double digesting test
- Digesting E0240 with EcoRI/PstI.
- E0240 Digestion system contains：
1 µl 10*H 0.25 µl EcoRI 0.25 µl PstI 5 µl Plasmid 3.5 µl dH20 -------------------------- 40 µl Total
electrophoresis result
- from left to right:
- E0240 -1 @ EcoRI/PstI
- E0240 -2 @ EcoRI/PstI
- pSB1A2
- Maker(DL2000 plus)
Double digesting test for PlacI-I741051 & R0010 & E0240
- use Double digesting test for gel extraction.
- PlacI-I74051 digestion system contains(Standard Assembly vector):
4 µl 10*H buffer 1 µl SpeI 1 µl PstI 20 µl Plasmid 14 µl dH20 -------------------------- 40 µl Total
- R0010 digestion system contains(Standard Assembly vector):
4 µl 10*H buffer 1 µl SpeI 1 µl PstI 20 µl Plasmid 14 µl dH20 -------------------------- 40 µl Total
- E0240 digestion system contains(Standard Assembly vector):
4 µl 10*M buffer 1 µl XbaI 1 µl PstI 20 µl Plasmid 14 µl dH20 -------------------------- 40 µl Total
electrophoresis result before gel extraction
- from left to right:
electrophoresis result after gel extraction
- from left to right:
