Lab Aug 14 2009

Came in at 8:30 and found the ccdB culture at an OD of .325 so I went ahead and prepped a new batch of competent cells following the top-10 protocol. Stored them in the -80 in an inner bag in the same bag where the old ccdB cells were.

Put the broth cultures in the fridge and turned off the water bath.

Put the plates in the fridge. Looks like we can pick the P1010 Cm and grow it from the plates as well(although may want to wait until Monday since otherwise the broth would be in all weekend)

1:00pm

Many parts miniprepped: Qiagen Miniprep Protocol

Ran two 1% agarose gels with the following parts: Agarose Gel Electrophoresis Protocol

Gel 1 - 2µL plasmid + 2µL 6x buffer - started (?? - assume 2:15pm), stopped 3:00pm

ladder 	Kan P1010 	p + RFP J23102 	Wintergreen J45120 	Banana J45200 	Quad K145303 	cI tripart P0151 	pAra I0500 B visible 	no band 	faint band 	band 	band 	faint band 	faint band 	no band

aug 14th gel 1.tif

Gel 2 - 2µL plasmid + 2µL 6x buffer + 8µL dH2O - started 3:05pm, stopped 3:50pm

ladder 	cI tripart P0151 	Kan P1010 	Aspartase C0083 	p + RFP J23102 	ECFP E0422 	p lambda cI R1051 	pAra I0500 A visible 	faint band 	no band 	band 	faint band 	faint band 	no band 	no band

aug 14th gel 2.tif

These results are somewhat as expected - we've been having problems with the P1010, the I0500, and the R1051//R0051 the entire time. Now I'm wondering if the P1010-Cm worked, either...

Cryopreserved : Cryopreservation Protocol

Wintergreen J45120, Banana J45200, Aspartase C0083, Kan P1010

--although, we belatedly realise that the P1010-Kan has no plasmid.

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