IGEM:Harvard/2006/Cyanobacteria/Notebook/2006-10-2

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Re-Digestions eg. peng is dumb

Previously, the B0034 should have been digested with \SP, and KaiC with \XP. Not sure what I was thinking there...

  • B0034 \SP + J36000 KaiC \XP
  • B0034 \SP + J36001 KaiB \XP

(Note: Just KaiA, B, or C on a backbone we can submit the geneart plasmids; as far as i can tell they do not have other E/X/S/P sites)

It looks like we have a signifigant amount of J36000 KaiC \XP.'

Then:


  • B0034 \SP: 8 lanes
  • J36001 \XP: 8 lanes


//Why 6? Fits perfectly into a 20 well gel!

Protocols:

B0034 \SP: 6 samples

    • 7.6 uL DNA (46.36)
    • 1 µL 10 x BSA (6.1)
    • 1 µL 10x buffer #2 (6.1)
    • 0.2 µL Spe1 (1.22)
    • 0.2 µL Pst1 (1.22)

J36001 \XP: 6 samples

    • 7.6 uL DNA (46.36)
    • 1 µL 10 x BSA (6.1)
    • 1 µL 10x buffer #3 (6.1)
    • 0.2 µL Xba1 (1.22)
    • 0.2 µL Pst1 (1.22)

The usual, @ 16h @45C.

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