Team:BYU Provo/Team Thermosensor/Week9

From 2011.igem.org

Team BYU Provo

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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.