IGEM:Peking/2007/Count-Conjugation-Notebook/2007-8-20: Difference between revisions
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*Preparation F+, R751 competent cell for Conjugation test. | *Preparation F+, R751 competent cell for Conjugation test. | ||
==mini-prep PlacI-I741051-E0240_pSB1A2, R0010-E0240_pSB1A2 (II) == | ==mini-prep PlacI-I741051-E0240_pSB1A2, R0010-E0240_pSB1A2 (II) == | ||
*last mini-prep digesting test didn't show the correct backbone.[[IGEM:Peking/2007/Count-Conjugation-Notebook/2007-8-18]] | *last mini-prep digesting test didn't show the correct backbone. | ||
*see:[[IGEM:Peking/2007/Count-Conjugation-Notebook/2007-8-18]] | |||
*Amplification Culture of PlacI-I741051-E0240_pSB1A2, R0010-E0240_pSB1A2 from plates. | *Amplification Culture of PlacI-I741051-E0240_pSB1A2, R0010-E0240_pSB1A2 from plates. | ||
*using Transgen mini plasmid puriflication kit. | *using Transgen mini plasmid puriflication kit. | ||
Revision as of 22:44, 20 August 2007
Tandem OriT by Qu Mingzhi & Ren Ze
Preparation of Competent Cells: F+, R751
- Preparation F+, R751 competent cell for Conjugation test.
mini-prep PlacI-I741051-E0240_pSB1A2, R0010-E0240_pSB1A2 (II)
- last mini-prep digesting test didn't show the correct backbone.
- see:IGEM:Peking/2007/Count-Conjugation-Notebook/2007-8-18
- Amplification Culture of PlacI-I741051-E0240_pSB1A2, R0010-E0240_pSB1A2 from plates.
- using Transgen mini plasmid puriflication kit.
- 50µL after purflication.
mini-prep double digesting test
- Digesting PlacI-I741051-E0240_pSB1A2, R0010-E0240_pSB1A2 with EcoRI/PstI.
- 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:
- 1 Maker(DL2000 plus)
- 2-3 PlacI-E0240 @ EcoRI/PstI
- 4-5 PlacI-I741051-E0240 @ EcoRI/PstI
