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Team:BYU Provo/Team Thermosensor/Week9
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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. | ||
</div> | </div> | ||
Latest revision as of 08:14, 24 September 2011
|
Contents |
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.
