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Ligation
From 2010.igem.org
(Difference between revisions)
(New page: 1. Mix together the following: a. 5 ul deionized water b. 2 ul of Reaction Buffer c. 5 ul of digested downstream insert (or water if using only 1 insert) d. 5 ul of dig...) |
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f. 1 ul of DNA ligase | f. 1 ul of DNA ligase | ||
| - | 2. Incubate at room-temp for | + | 2. Incubate at room-temp for 5 mins |
3. Kill reaction for 20 mins at 80 degrees Celsius | 3. Kill reaction for 20 mins at 80 degrees Celsius | ||
Latest revision as of 15:17, 29 July 2010
1. Mix together the following:
a. 5 ul deionized water
b. 2 ul of Reaction Buffer
c. 5 ul of digested downstream insert (or water if using only 1 insert)
d. 5 ul of digested downstream insert
e. 3 ul of digested vector
f. 1 ul of DNA ligase
2. Incubate at room-temp for 5 mins
3. Kill reaction for 20 mins at 80 degrees Celsius
