User:Karmella Haynes/Notebook/BioBrick cloning/2014/12/23

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Karmella's BioBrick Cloning Main project page
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12/23/14

  • Planning: Gal4 fusion cloning



Planning: Gal4 fusion cloning

  • "Backbone" = Gal4-mCherry-VP64 construct KAH60/pcVN
  • Approach: swap-in activator domain to replace VP64
    • PCR-amplify backbone (omitting VP64)
    • PCR-amplify insert
    • Plan A: use LCR to ligate insert into backbone


Order Primers (01/02/15)
Attempt 1: swap-in ATF activation domain; add 5' phosphates to all oligos

  • KAH60 "backbone" (without VP64, includes scars) = 6637
    • KAH60 F1
    • KAH60 R1
  • ATF2 insert = 906
    • ATF2_f1
    • ATF2_r1
  • LCR Bridging oligos
    • LCRb_KAH60_ATF2_1
    • LCRb_KAH60_ATF2_2





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