Team:BYU Provo/Team Thermosensor/Week6


Team BYU Provo

BYU Provo

Week 6 (May 22 - 28)

  • May 23 LacZ ran on gel-product YES!
    • LacZ PCR cleanup
      • Cleaned up LacZ according to Qiaquick protocol and labeled as T 20-1(needs to be checked on gel)
  • May 25
    • Restriction Digest of LacZ and GFP
      • Digested GFP at the HindIII and XbaI sites
        • oops...we realized that our reverse LacZ primer had a BglII site in it when we needed an XbaI site. Dr. Grose ordered a new primer with an XbaI restriction site in it.
    • Confirming pBAD clones by restriction digest
      • 5uL buffer 4
      • 0.5 uL BSA
      • 30 uL plasmid(pIG 17-4 and 17-5 each in individual tubes)
      • 12uL H2O
      • 1.5 uL BamHI-HF
      • 1.5 uL EcoRI-HF
        • If the gel shows two bands then pBAD was inserted successfully into pIG17-4 and/or pIG17-5. The Digest of pIG174 will be labeled as RD 20-4 and the digest of pIG17-5 will be labeled RD 20-5.
        • placed RDs in 37oC bath overnight and placed in the freezer
  • May 27
    • Running RD 20-4 and RD 20-5 on an agarose gel. We will be using a digested pBAD gene and pPLAT digested at BamHI and PstI as our controls on the gel.