Freiburg07/Transformation

From 2007.igem.org

(Difference between revisions)
 
(3 intermediate revisions not shown)
Line 1: Line 1:
-
Transformation of frozen competent cells
+
'''Transformation of frozen chem. competent cells'''
-
1. Thaw frozen cells on ice, 10-15 minutes.
+
1. Thaw frozen cells.<BR>
-
2. Add 80 µl cells to the 20 µl ligation reaction.
+
2. Add 10 µl ligation and shake gently.<BR>
-
3. Incubate on ice for 5 minutes.
+
3. Incubate on ice for 5-10 minutes.<br>
-
4. Heat shock cells for 10 minutes at 37 °C.
+
4. Heat shock cells for 10-20 minutes at 37 °C.<BR>
-
5. Bring up in 1 mL LB and shake gently for 1 hour at 37 °C.
+
5. Add 900 µl DYT-Medium and shake gently for 60-70 minutes at 37 °C.<br>
 +
 
 +
[https://2007.igem.org/Freiburg BACK]

Latest revision as of 15:00, 24 October 2007

Transformation of frozen chem. competent cells

1. Thaw frozen cells.

2. Add 10 µl ligation and shake gently.

3. Incubate on ice for 5-10 minutes.

4. Heat shock cells for 10-20 minutes at 37 °C.

5. Add 900 µl DYT-Medium and shake gently for 60-70 minutes at 37 °C.

BACK