"
USTC/Notebook/DNA ligation
From 2008.igem.org
(Difference between revisions)
| Line 7: | Line 7: | ||
Materials | Materials | ||
| - | *'''DNA plasmids'''; . Measure concentration in Nanodrop beforehand. | + | *'''DNA plasmids''' double digested previously(eg.XbaI and PstI); . Measure concentration in Nanodrop beforehand. |
| - | *'''Biobricks''' | + | *'''Biobricks''' double digested previously(eg.SpeI and PstI ) |
*equal amount of T4 ligase. | *equal amount of T4 ligase. | ||
Revision as of 11:55, 28 October 2008
DNA Ligation
DNA ligases are used with restriction enzymes to insert genes into plasmids.
Materials
- DNA plasmids double digested previously(eg.XbaI and PstI); . Measure concentration in Nanodrop beforehand.
- Biobricks double digested previously(eg.SpeI and PstI )
- 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.
