Team:BYU Provo/Team Thermosensor/Week9


Team BYU Provo

BYU Provo


Week 9 (June 12-18)

14 June 2011

Dr. Grose approved our pBAD insert, just said to run a control next time on the gel. Set up overnight on colonies from T26-1 lig plate from slices 2 and 5 (like a pizza! Insert drawing of a pizza)

  • T28-2-1 through T28-2-4
  • 4 ml LB broth
  • 500 ul Amp
  • 1 tip dipped in colony from slice 2
  • T28-5-1 through T28-5-4
  • 4 mL LB broth
  • 500 ul Amp
  • 1 dipped in colony from slice 5
    • Placed in 37*C shaker @ 10:41 AM

15 June 2011

Placed T28-2 through 4 and T28-5-1 through 4 in 4* “incubator” until someone can due a miniprep on them.

16 June 2011

Began Qiaprep Miniprep purification protocol on tubes T28-2(1-4) and T28-5(2-4)

  • MP T29-2-1 = T28-2 (1+4)
  • MP T29-5-1 = T28-5 (2+4)
  • MP T29-2-2 = T28-2 (2+3)
  • MPT29-5-2 = T28-5 (1+3)
    • all of T29-2 was combined into one and all of T29-5 was combined into one
  • Colony 2 plasmid is pIG12
  • Colony 5 plasmid is pIG13
  • We need to find the concentration of both of these plasmids with the nanodrop machine, then we can due a restriction digest with lacZ and GFP and insert both of them
  • pIG 12 -> 137.8 ng/ul
  • pIG 13 -> 127.8 ng/ul

17 June 2011

We did a Restriction Digest on our pLATBAD construct (pIG12) as well as our LacZ product and GFP product. They’ll be inserted at our HindIII and XbaI sites. Started incubating at 2:15pm and removed from the 37oC incubator at 3:50pm. We then ran a low melt gel on the two inserts and pLATBAD at 90V for 45 min. Placed low melt in fridge. Lets check the RD with Dr. Grose before continuing.