8.30am
Both plates from last night have colonies. (E0840, and K115020).
Cleaned miniprep columns. Cleaning miniprep spin columns
Started miniprep.
1.30pm
Prepared gel.
4pm
Gel worked! Everything was there except for R0051 which was missing all three times.
Oct 10 gel 1.tif
Ladder |
K235019A |
K235020A |
K235021A |
K115017A |
K115017D |
C0012A |
J04630A |
J23070A |
R0051A |
K235019B |
K235020B |
K235021B |
K115017B |
K115017E |
C0012B |
J04630B |
J23070B |
R0051B |
Blank |
Band? |
Band |
Band |
Band |
Band |
Band |
Band |
Band |
Band |
NO |
Band |
Band |
Band |
Band |
Band |
Band |
Band |
Band |
NO |
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Comments |
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Ladder |
K235019C |
K235020C |
K235021C |
K115017C |
K115017F |
C0012C |
J04630C |
J23070C |
R0051C |
Band? |
Band |
Band |
Band |
Band |
Band |
Band |
Band |
Band |
NO |
Comments |
New miniprep column |
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New miniprep column |
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New miniprep column |
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New miniprep column |
This also shows that our miniprep cleaning is working as well, as the new and reused ones look indistinguishable.
I think the biggest issue for these is having a growth time of exactly 12-16 hours, and miniprepping immediately at that point. I'm guessing that putting them in the fridge was our big problem.
We don't have enough epi's to do cryopreserve stocks however.
I made broth cultures from E0840, and K115020. I'll be back tomorrow at 8.30am to miniprep them.
10pm
Started assemblies K235030 Kan, K235030 Chl, and K235031 Chl.
K235030 Kan: J23102(Amp) const. promoter + K115017(Amp/Tet) 32oC temp. sensing RBS + P1010 Kan
K235030 Chl: SAME + P1010 Chl
K235031 Chl: C0012(Amp) LacI repressor+LVA + B0015(Amp/Kan) transcription stop + P1010 Chl
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