Team:Gaston Day School/17 June 2009

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(New page: <html> <du> <dt>High Efficiency E. Coli</dt> <dt>Thaw tube of NEB 10-beta Comp. E. Coli cells on ice until ice disappears. Gently mix and pipette 50mL into transformation tube on ice.</dt>...)
 
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<dt>High Efficiency E. Coli</dt>
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<dt>High Efficiency E. coli</dt>
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<dt>Thaw tube of NEB 10-beta Comp. E. Coli cells on ice until ice disappears. Gently mix and pipette 50mL into transformation tube on ice.</dt>
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<dt>Thaw tube of NEB 10-beta Comp. E. coli cells on ice until ice disappears. Gently mix and pipette 50mL into transformation tube on ice.</dt>
<dt>Add plasma DNA and carefully mix</dt>
<dt>Add plasma DNA and carefully mix</dt>
<dt>Suspend the well 8E on Plate 2 in 15x TE</dt>
<dt>Suspend the well 8E on Plate 2 in 15x TE</dt>

Latest revision as of 14:24, 20 July 2009

High Efficiency E. coli
Thaw tube of NEB 10-beta Comp. E. coli cells on ice until ice disappears. Gently mix and pipette 50mL into transformation tube on ice.
Add plasma DNA and carefully mix
Suspend the well 8E on Plate 2 in 15x TE
Add 5x of Plasmid to E. Coli
Place on ice for 30 mins
Heat shock at 42C for 30 seconds
Ice for 5 mins
Pipette 950microliters of room temp SOC
Place 37C for 60 mins and shake every 15 mins
Warm plates to 37C
Spin 5 mins
Pipette out SOC
Resuspend in 150microliters SOC
Mix tubes
Spread 50-100microliters onto plate and incubate overnight at 37-42C
Plate 1 = 100microliters
Plate 2 = 50microliters

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