Summary
- Digest the pSB1A2/3 plasmid (from Rene) with XbaI & SpeI to get rid of the insert
Reagent |
Volume |
|
|
DNA (plasmid) |
20.0
|
10x buffer |
3.0
|
XbaI |
1.0
|
SpeI |
1.0
|
dH2O |
5.0
|
|
30 μL --> 37°C/ ~10 min.
|
- Run entire 30 μL on a 1% gel (use big tooth comb) next to a ladder
- You should see two bands. The larger band (2000 bp) is the backbone.
- Cut out and gel purify this fragment (ignore the shorter fragment).
- Notify Dr. Haynes when you are done. The next step is Gibson Assembly and transformation.
|