Wisconsin-Madison/26 May 2009

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