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From 2010.igem.org

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Revision as of 19:14, 27 October 2010

10/20/10

Results for plating

Plate B showed a white colony growth

Plates A, C, D have shown no signs of growth for 1 day

Keeping all plateed in 37C incubator for additional day

Joe and I did a second restriction assay protocol

I performed the steps exaction line the previous assay

Joe performed the steps for 10 ul of LacZ and 10 ul of T9002

I performed the steps for 30 ul of LacZ and 30 ul of T9002

Enzyme Parts

NE Bufferx, Biolabs, # B7002S, Lot# 0030910

Pruified BSA, Biolabs, #B9001S, Lot# 0101002

Eco RI-HF, Biolabs #R3101S, Lot# 0021002

Xbai, Biolabs, #R0145S, Lot#038000912

Spe I, Biolabs, #R0133S, Lot# 0J10911

I spun down all four tubes containing LacZ and T9002

I placed them in the thermocycler 30 min. @ 37C, then 80C for 20 min

Cut and ligation of Lac Z to T9002 plasmid 10/18/10

combination of digests 

Notes:

set thermocycler to 16 ul for ligation

start 6:26 finished 7:18 no exceptions

Transformation of NEB E.coli 10/15/10

5 ul each E.coli cuvette

cescue w/SOB

spread on galx/Amp plates