Difference between revisions of "Part:BBa K649200:Experience"
| Line 36: | Line 36: | ||
'''[Method]''' | '''[Method]''' | ||
<br /> | <br /> | ||
| − | 1. The part on pSB1C3 was made linear by EcoRV restriction site which is far from lox sites. <br /> | + | 1. The part on pSB1C3 was made linear by EcoRV restriction site which is far from lox sites. <br /><br /> |
| − | 2. Three identical samples and one negative control without Cre-recombinase had been prepared and those were incubated in 37°C for 0.5hr, 2hr and 4hr respectively. Negative control was incubated for 4hr.<br /> | + | 2. Three identical samples and one negative control without Cre-recombinase had been prepared and those were incubated in 37°C for 0.5hr, 2hr and 4hr respectively. Negative control was incubated for 4hr.<br /><br /> |
3. After a period of time, the samples were kept in -20°C until the whole samples are arranged. The result was checked by electrophoresis(0.8% EtBr(+) agarose gell, 100V, 400mA, 30min) | 3. After a period of time, the samples were kept in -20°C until the whole samples are arranged. The result was checked by electrophoresis(0.8% EtBr(+) agarose gell, 100V, 400mA, 30min) | ||
Revision as of 08:42, 4 October 2011
Applications of BBa_K649200
In in vitro assay, the part was made linear using restriction enzymes. Cre recombinase was added to the linear DNA and incubated for 0.5, 2, and 4 hours. Images of the experiments have been added below.
[Sample]
| sample | volume |
|---|---|
| PlacIQ-lox2272-GFP-lox2272(pSB1C3) | 50 ng/μl × 4 μl |
| 10× Cre recombination Buffer | 1 μl |
| ddH2O | 4 μl |
| Cre recombinase | 1,000 units/ml × 1 μl |
| total | 10 μl |
[Method]
1. The part on pSB1C3 was made linear by EcoRV restriction site which is far from lox sites.
2. Three identical samples and one negative control without Cre-recombinase had been prepared and those were incubated in 37°C for 0.5hr, 2hr and 4hr respectively. Negative control was incubated for 4hr.
3. After a period of time, the samples were kept in -20°C until the whole samples are arranged. The result was checked by electrophoresis(0.8% EtBr(+) agarose gell, 100V, 400mA, 30min)
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