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Team:HokkaidoU Japan/Notebook/aggregation Week 5
From 2012.igem.org
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Now, I think like this.<br /> | Now, I think like this.<br /> | ||
The E. coli which we use transformation is BL21(DE3)pLysS. | The E. coli which we use transformation is BL21(DE3)pLysS. | ||
| - | So, the E. coli could multiplicationincrease on LBC plate.< | + | So, the E. coli could multiplicationincrease on LBC plate.<br /> |
I'm going to do transformation , use DH5α. | I'm going to do transformation , use DH5α. | ||
Revision as of 13:05, 30 July 2012
July 30th
digestion
I tried to digeste the mini-prep products again, and after Ethanol precipitation, we use electrophoresis.
transformation
Now, I think like this.
The E. coli which we use transformation is BL21(DE3)pLysS.
So, the E. coli could multiplicationincrease on LBC plate.
I'm going to do transformation , use DH5α.
Transformation of plasmid DNA ligated in 25th (pT7-RBS-Ag43-dT on pSB1K3) in E. coli(DH5α).
- Added 2ul of plasmid DNA to 50ul of thawed competent cells on ice.
- Incubated on ice for 30min.
- Added 200ul of LB then incubated the cells for 2 hours at 37C.
- Prepared and Labeled two petri dishes with LBK.
- Plate 200ul of the transformation onto first dish and spread.
- Added 450ul of LB to 50ul of the transformation and plated 200ul of it onto second dish and spread.
- Incubated the plates at 37C for over 30hrs.
