User:Meng Xiao He/Notebook/fall08/2008/10/31

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Update

  • BsmBI digest did not over digest (band sizes still correct)
  • T4 ligation for 15 minutes made DNA smear into large (8-10Kb) region, so over ligated

Next

  • repeat
  • PCR fragments over using existing fragments
    • purify>dephosphorylate fragments>purify>digest with BsmBI>ligate for 1-5 minutes>purify>XmaI digest
  • try crossover PCR

PCRs:

1-6:

  • cbb, cco, Duo; 3 then 5

Xover:

  • cbb, cco, Duo