Team:Exeter/lab book/1gp/wk2

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ExiGEM2012 Lab Book 1GP wk2

Single Gene Plasmids and Enzyme Characterisation: 16th - 20th July 2012

Monday 16th July (11.00)

BioBrick extraction of RBS BioBrick (BBa_B0034) and TetR repressible promoter (BBa_R0040)

Transformation of RBS BioBrick and TetR promoter

Tuesday 17th July (15.00)

Adding cultures with RBS and TetR promoter into liquid medium and incubation overnight

Protocol was followed using ampicillin for all BioBrick parts as the selection antibiotic.

Wednesday 18th July (15.00)

Adding cultures with RBS and TetR promoter into liquid medium and incubation overnight

No cultures were found in all liquid mediums and thus repeated.

Thursday 18th July (9.00)

Mini-Prepping of RBS and TetR promoter

• Gel Electrophoresis to check fragment sizes of RBS and TetR promoter

Gel Electrophoresis showed that RBS and TetR promoter were successfully cloned. Lane 1 = DNA hyperladder, Lane 3 = TetR promoter mini-prep 2 (expected: 2155bp since TetR promoter is too small to be seen at 54bp), Lane 4 = TetR promoter mini-prep 3 (expected: 2155bp since TetR promoter is too small to be seen at 54bp), Lane 5 = TetR promoter mini-prep 4 (expected: 2155bp since TetR promoter is too small to be seen at 54bp), Lane 6 = RBS mini-prep 1 (expected: 2155bp since RBS is too small to be seen at 12bp), Lane 7 = RBS mini-prep 2 (expected: 2155bp since RBS is too small to be seen at 12bp), Lane 8 = RBS mini-prep 3 (expected: 2155bp since RBS is too small to be seen at 12bp), Lane 9 = RBS mini-prep 4 (expected: 2155bp since RBS is too small to be seen at 12bp), Lane 10 = DNA hyperladder.

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