Virginia Commonwealth/27 May 2009
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==Wednesday 27 May 2009== | ==Wednesday 27 May 2009== | ||
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===Tasks=== | ===Tasks=== | ||
- | # | + | # Transform NEB10beta with J04450 and I13521, using frozen stock of comp. cells. |
- | # | + | |
+ | ===Results=== | ||
+ | # Kept all tubes and cuvettes on ice at all times, and thawed one tube of NEB10beta competent cells (250uL). Each of the following transformation was performed individually: | ||
+ | #* (-) control, with no DNA added | ||
+ | #* 1uL J04450 and 50uL cells | ||
+ | #* 2uL J04450 and 50uL cells | ||
+ | #* 1uL I13521 and 50uL cells | ||
+ | # Added 50uL cells to cold microcentrifuge tube. | ||
+ | # To cells, added appropriate volume and type of DNA, mixing well by pipette, and returning to ice to incubate for 1min. | ||
+ | # Transfer mix to cold 2mm electrotransformation cuvette, tapping to get mixture to the bottom. | ||
+ | # Pulsed cuvette on Ec2 setting, then immediately added 1mL LB. | ||
+ | # Transferred mixture to 15ml centrifuge tube and incubated for 30min at 37*C. | ||
+ | # Spread 35uL on either kan (J04450 and ctrl) or amp (I13521), incubated at 37*C overnight. |
Latest revision as of 03:38, 28 May 2009
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Wednesday 27 May 2009
Tasks
- Transform NEB10beta with J04450 and I13521, using frozen stock of comp. cells.
Results
- Kept all tubes and cuvettes on ice at all times, and thawed one tube of NEB10beta competent cells (250uL). Each of the following transformation was performed individually:
- (-) control, with no DNA added
- 1uL J04450 and 50uL cells
- 2uL J04450 and 50uL cells
- 1uL I13521 and 50uL cells
- Added 50uL cells to cold microcentrifuge tube.
- To cells, added appropriate volume and type of DNA, mixing well by pipette, and returning to ice to incubate for 1min.
- Transfer mix to cold 2mm electrotransformation cuvette, tapping to get mixture to the bottom.
- Pulsed cuvette on Ec2 setting, then immediately added 1mL LB.
- Transferred mixture to 15ml centrifuge tube and incubated for 30min at 37*C.
- Spread 35uL on either kan (J04450 and ctrl) or amp (I13521), incubated at 37*C overnight.