User:Meng Xiao He/Notebook/fall08/2008/11/06

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pir-116 maintains plasmids at high copy

  • higher than glycerol stock strain, 300-400 ng/ul

Distinguishing digests

  • TOPO (3.5kb): EcoRI+PstI
    • Duo: 1.2 kb, 300bp
    • cco: 1.7 kb
    • cbb3: 400, 830, 300
  • pER21+flanks (and flankless as ctrl):
    • EcoRI + PstI>column>SmaI at 25

Gel 1: pER21+flanks, EP>SmaI digest

Image:11.6.EPS_digest1.mx.jpg

  • 1-3: A1-3
  • 100 bp ladder
  • 5-7: B1-3
  • pER21
  • 1kb ladder
  • 10-12: C1-3

B2 is cbb3 candidate & C1 is cco candidate

Gel 2

Image:11.6.EPS_digest2.mx.jpg

  • pER21: EP only
  • 1kb ladder
  • 3-5: TOPO A, B, C
  • 100 bp ladder
  • A1-3 EP only
  • B1-3 EP only
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