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Team:Brown/Notebook/September15
From 2010.igem.org
Wednesday, September 15, 2010
Restriction digests
1) J13002 (TetR binding sites + RBS) with Spe1, Pst1 2) LacI A (RFC25) with Xba, PstI 3) AraC (RFC25) with Xba, PstI
1) 50ul total volume
- 5ul NEB buffer 2
- 3ul DNA (J13002)
- 0.5ul 100x BSA
- 1ul SpeI
- 1ul PstI
- 41.5ul dH2O
2) 50ul total volume
- 5ul NEB buffer 3
- 3ul DNA (LacI)
- 0.5ul 100x BSA
- 1ul SpeI
- 1ul PstI
- 41.5ul dH2O
3) 50ul total volume
- 5ul NEB buffer 2
- 3ul DNA (AraC)
- 0.5ul 100x BSA
- 1ul SpeI
- 1ul PstI
- 41.5ul dH2O
37C for one hour, then put in freezer.
Notes - NEB Double Digest Finder: PstI SpeI -> Buffer 2. PstI Xba -> buffer 3
This is not an ideal suggestion, buffer/enzymes are not 100% compatible
