Team:Osaka/Protocols

(Difference between revisions)

Revision as of 07:36, 24 October 2010

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.
SOC medium
- Add sterilized 2M glucose 1 / 100 volume to SOB medium, and sterilize by filtration using 0.22 um microfilter. Store at 4℃.
Tranformation buffer, TB 1L(-> final conc.)
- PIPES 3.0g(-> 10mM)
- CaCl2 2H2O 2.2g(-> 15mM)
- KCl 18.6g(-> 250mM)

@@ Line 7: / Line 7: @@
 *Preparation of competent cells for heat-shock transformation
 ** Preparation
-# SOB medium            1L(-> final conc.)
+# SOB medium 1L(-> final conc.)
-#* Bacto Tryptone       20g(-> 2.0%)
+#* Bacto Tryptone 20g(-> 2.0%)
-#* Bacto Yeast Extract  5g(-> 0.5%)
+#* Bacto Yeast Extract 5g(-> 0.5%)
-#* 5M NaCl              2ml(-> 10mM)
+#* 5M NaCl 2ml(-> 10mM)
-#* 2M KCl               1.25ml(-> 2.5mM)
+#* 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.
+# SOC medium
+#* Add sterilized 2M glucose 1 / 100 volume  to SOB medium, and sterilize by filtration using 0.22 um microfilter. Store at 4℃.
+# Tranformation buffer, TB 1L(-> final conc.)
+#* PIPES 3.0g(-> 10mM)
+#* CaCl2 2H2O 2.2g(-> 15mM)
+#* KCl 18.6g(-> 250mM)
 *Transformation by heat-shock
 *Plasmid DNA miniprep