USTC/Notebook/DNA ligation
From 2008.igem.org
(Difference between revisions)
(New page: '''< Back to Notebook''' == DNA Ligation ==) |
|||
Line 2: | Line 2: | ||
== DNA Ligation == | == DNA Ligation == | ||
+ | |||
+ | DNA ligases are used with restriction enzymes to insert genes into plasmids. | ||
+ | |||
+ | |||
+ | Materials | ||
+ | *'''DNA plasmids'''; . Measure concentration in Nanodrop beforehand. | ||
+ | *'''Biobricks''' | ||
+ | *equal amount of T4 ligase. | ||
+ | |||
+ | |||
+ | |||
+ | Procedure | ||
+ | |||
+ | The following volumes apply to a 20µl DNA ligation. | ||
+ | |||
+ | add the following components to PCR tube: | ||
+ | *'''9µl''' DNA plasmids | ||
+ | *'''1µl''' biobricks | ||
+ | *'''10ul''' T4 ligase | ||
+ | *'''MIX''' the reaction by stiring | ||
+ | *Incubate the reaction at 16*C for 2hrs or overnight to ensure complete ligation. |
Revision as of 11:50, 28 October 2008
DNA Ligation
DNA ligases are used with restriction enzymes to insert genes into plasmids.
Materials
- DNA plasmids; . Measure concentration in Nanodrop beforehand.
- Biobricks
- equal amount of T4 ligase.
Procedure
The following volumes apply to a 20µl DNA ligation.
add the following components to PCR tube:
- 9µl DNA plasmids
- 1µl biobricks
- 10ul T4 ligase
- MIX the reaction by stiring
- Incubate the reaction at 16*C for 2hrs or overnight to ensure complete ligation.