Team:Wash U/Protocol
From 2009.igem.org
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:8. Label two petri dishes with LB agar and the appropriate antibiotic(s) with the part number, plasmid, and antibiotic resistance. Plate 20 µl and 200 µl of the transformation onto the dishes, and spread. This helps ensure that you will be able to pick out a single colony.<br> | :8. Label two petri dishes with LB agar and the appropriate antibiotic(s) with the part number, plasmid, and antibiotic resistance. Plate 20 µl and 200 µl of the transformation onto the dishes, and spread. This helps ensure that you will be able to pick out a single colony.<br> | ||
:9. Incubate the plate at 37ºC for 12-14 hours, making sure the agar side of the plate is up. If incubated for too long the antibiotics start to break down and un-transformed cells will begin to grow. This is especially true for ampicillin because the resistance enzyme is excreted by the bacteria, and inactivate the antibiotic outside of the bacteria.<br> | :9. Incubate the plate at 37ºC for 12-14 hours, making sure the agar side of the plate is up. If incubated for too long the antibiotics start to break down and un-transformed cells will begin to grow. This is especially true for ampicillin because the resistance enzyme is excreted by the bacteria, and inactivate the antibiotic outside of the bacteria.<br> | ||
+ | :Note: Restriction sites E=EcoR1-HF; X=Xba1; S=Spe1; P=Pst1; M=Mixed Site | ||
:To view the full BioBrick Manual procedures, please click [http://partsregistry.org/Help:Transformation_Protocol here]. | :To view the full BioBrick Manual procedures, please click [http://partsregistry.org/Help:Transformation_Protocol here]. | ||
Revision as of 17:11, 30 June 2009