Lab Aug 12 2009

Last night's plates:

E0422 - ECFP tripart - good growth

K145303 - GFP with ribolock - good growth

J23102 - constitutive promoter - good growth (red colonies!)

I0500 - pAra - no growth

R1051 - p-lambda cI - one isolated, suspiciously small colony

broth culture the 3 parts that grew. (not culturing K145303, just putting it in the fridge. We have a broth culture from the previous day's plate)

Miniprep K145303, P0151 (waiting until thursday when we have more parts)

plated a Kan plate with a known good Kan resistant cell line (from Adrienne)

cryopreserve our DH5alpha cells (grown back from plate before the plate got too dry, just to keep our cell line...) Also preserve P0151, K145303.

rehydrate and transform:

K081005 - constitutive promoter plus strong RBS

pSB1A2, plate 2, well 10F

C0083 - aspartase

pSB2K3, plate 2, well 17A

J45200 - banana

pSB1AT3, plate 2, well 5F

J45120 - wintergreen

pSB1AT3, plate 2, well 5D

K098988 - temperature sensitive λcI-controlled GFP generator

pSB3K3, plate 3, well 11H

Transform P1010 Kan and Cm backbones into competent DB3.1 cells (with pUC as control)

-We used 200ul competent cells instead of the normal 100ul to compensate for our diluting the cells with 40% Glycerol.

Transform new rehydations from today

-All transformed parts went into final incubation step at 2:10pm

-Amp plasmids (J45200, J45120, K081005) come out for plating at 3:25pm

-Kan plasmids (C0083, P1010 Kan and K098988) come out for plating at 4:10pm

-Cm plasmids (P1010 Cm) come out for plating at 4:10pm

-pUC18 plasmid come out for plating at 3:25pm and 4:10pm, to see if extra transformation time works better for the DB3.1 cells

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