Kyliah says: I'll come in today for the afternoon. Did the stabs ever show up? If not, we might be able to substitute something out for the arabinose operon.
11.00am
Well nothing grew last night so we have some problems. There is one tiny suspicious colony on a P1010 kan plate but I can't imagine it will be correct.
On the plus side, between Dr. Upton and Dr. Nano we should have bench space, along with 4C, -20C and -80C space, so everything we need for the fall!
I'm going to start by running a gel for the parts used yesterday: J23032(Amp) downstream, J23102(Amp) upstream, B0015(AmpKan) downstream, J06504(Amp) upstream, K235000(Cm) downstream
Gel 1 September 1
Started at 2.30pm
|
1 Kb Ladder
|
J23032 Down |
J23102
Up
|
B0015 Down |
J06504 Up |
K235000 Down |
P K081005 |
P K098988 |
| |
No |
Band |
Band |
Band |
No |
No |
Band |
sep 1 gel 1.tif
K235009=J23102(Amp)+J23032(Amp)+P1010Cm - No wonder this assembly didn't work, one of the ligation parts seems to be absent
K235010=J06504(Am)+B0015(AmpKan)+P1010Cm - This one should have worked, though.
(back to Lab notebook)
Comments (1)
Layne Woodfin said
at 10:48 pm on Sep 2, 2009
I was advised by one of the TAs and a professor with a fair bit of experience with gels that parts under 100bp won't show up on an ethidium gel, this might explain the absence of the parts K081005 and J23032 as they're under 100bp (the plasmid still should have shown up though... so maybe there's just nothing there as was suggested). For parts under 100bp, we're going to need another way to verify their presence. Also, after looking at the gel for some time I'm convinced that there's a faint band in lane 5 (J235000)
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