Lab Oct 10 2009

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

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