Purdue/2 September 2008

From 2008.igem.org

Revision as of 17:05, 2 September 2008 by Mstine (Talk | contribs)
(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)

Click Here to return to the notebook.

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

Retrieved from "http://2008.igem.org/Purdue/2_September_2008"