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Purdue/2 September 2008
From 2008.igem.org
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Restriction Digest of SOS and LacZ
1. Mixed digestion solution:
- 50uL DNA
- 7uL NEB2
- 10.5uL ddH2O
- 0.5uL BSA
- 1uL of each enzyme
- SOS: EcoRI + SpeI
- LacZ: EcoRI + XbaI
- This makes SOS the small part cut out (192 bp) and put into the LacZ plasmid (~5000 bp...=LacZ + most of plasmid)
2. Put mixtures at 37C for 1 hour, 25 min.
3. Run through Agarose gel w/ DNA ladder, DNA loading dye (Bio-RAD)
- Mixed 7uL of loading dye w/ each 70uL tube
- 10uL ladder in lane 1
- Lanes 2-4: 20uL LacZ in each lane
- Lanes 5-7: SOS
- 120V, 45 min.
4. Overnight stain w/ Fast Blast
- 5x
- RT
- dark
- rocking
Edited by Janie Stine
