Wisconsin-Madison/26 May 2009
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May 26, 2009
Ex 3: Tested competent cells with pTrc99A which has Amp resistance
Ex1,2: Miniprep of NudF accoding to miniprep protocol
Ran gel with restriction digest (BAM H1 for 99, F,R) (EcoR5 for 18, 33) according to Restriction digest protocol
Results: Gel 1: Messed up first gel, Misplaced Ladder, Ran a second one:
Gel 1:
Gel 2:
Expectations:
pBAD33: 5352 bp (EcoR5)
pBt2BAD18: 4217 bp (EcoR5)
pTrc99A: 4176 bp (BAM H1)
NudF: 4716 bp (BAM H1)
YhfR: 4740 bp (BAM H1)
Results: Gel 2: correct plasmids and coding sequences, need better clean-up of RNA
- tested competent cells w/ pTrc99A on Amp50 plates (results show poor competency, only 150 colonies on undiluted plate) - NP
- mini preparation of pNudF (successful) - NP
- ran gels with digested plasmids to test length, verify plasmids (successful) - NP