User:Meng Xiao He/Notebook/fall08/2008/11/01

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PCR results

11.1.mx.xover.jpg

Lanes

  • 1-6:
    • cbb3: 3, 5; cco: 3, 5; Duo: 3, 5
  • 100bp ladder
  • 8-10 Xover:
    • cbb3, cco, Duo
  • 1kb ladder

X over product PCR

Since X over products were not very concentrated and had other amplicons, set up 25ul PCRs of just those 3 using some purified product as the template. PCR conditions: MXFG program, column 5, extension time changed to 45s.

  • This PCR ended up being very messy, with lots of smearing. There also appeared to be amplication of the fragments that made up the x-over individually. ??

XmaI o/n digest

  • cbb, cco, Duo from Xover PCR (original) [45, 70, 60 ul of DNA digested+ 2 of enzyme and appropriate vol of NEB4]
  • pUC19
  • pER21 old, concentrated old, and new
    • vectors were 20ul DNA+1 of enzyme and necessary NEB4