"
Team:Osaka/Protocols
From 2010.igem.org
(Difference between revisions)
Kagatataku (Talk | contribs) (→Protocols) |
Kagatataku (Talk | contribs) (→Protocols) |
||
| Line 6: | Line 6: | ||
*Preparation of competent cells for heat-shock transformation | *Preparation of competent cells for heat-shock transformation | ||
| - | **Preparation | + | ** Preparation |
| - | + | # SOB medium 1L(-> final conc.) | |
| + | #* Bacto Tryptone 20g(-> 2.0%) | ||
| + | #* Bacto Yeast Extract 5g(-> 0.5%) | ||
| + | #* 5M NaCl 2ml(-> 10mM) | ||
| + | #* 2M KCl 1.25ml(-> 2.5mM) | ||
| + | #* Add miliQ water 990ml to all miture and sterilize by autoclave. Before using, add sterilized Mg2 solution(1M MgSO4, MgCl2) 10 ml. | ||
*Transformation by heat-shock | *Transformation by heat-shock | ||
Revision as of 07:23, 24 October 2010
Protocols
- Preparation of competent cells for heat-shock transformation
- Preparation
- SOB medium 1L(-> final conc.)
- Bacto Tryptone 20g(-> 2.0%)
- Bacto Yeast Extract 5g(-> 0.5%)
- 5M NaCl 2ml(-> 10mM)
- 2M KCl 1.25ml(-> 2.5mM)
- Add miliQ water 990ml to all miture and sterilize by autoclave. Before using, add sterilized Mg2 solution(1M MgSO4, MgCl2) 10 ml.
- Transformation by heat-shock
- Plasmid DNA miniprep
- 3A BioBrick assembly
- PCR cloning
- PCR site-directed mutagenesis
- Extraction of genome DNA for PCR cloning
