"
Team:Brown/Notebook/August16
From 2010.igem.org
Monday, August 16, 2010
Miniprep of CI-pGEM (from XL1B)
- CI - 314ng/ul
Double digest of Mnt-pGEM, Lovtwo-pGEM
50ul total volume
- 5ul NEB Buffer 3
- 2ul DNA
- 0.5ul 100x BSA
- 1ul BglII
- 1ul XhoI
- 40.5ul dH2O
Gel of the above digest
- Ladder
- Lovtwo
- Mnt
Re-ligations
We are using more insert because the previous ligations were not fully successful.
- Lovtwo into pGEM
- bgl-CI into pGEM
- bgl-AraC into pGEM
- bgl-Mnt into pGEM
Concentrations:
- pGEM - 50ng/ul
- bgl-CI - 17.3ng/ul
- bgl-AraC - 14.2ng/ul
- bgl-Mnt - 15ng/ul
17ul total volume:
- 7.5ul ligase buffer
- 1ul T4 DNA ligase
- 0.5ul pGEM vector
- 8ul DNA
