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Lab Oct 10 2009
From 2009.igem.org
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
Comments
Ladder K235019C K235020C K235021C K115017C K115017F C0012C J04630C J23070C R0051C Band? Band Band Band Band Band Band Band Band NO Comments New miniprep column New miniprep column New miniprep column 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
