Team:Brown/Notebook/Protocols
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#Check Buffers P2 and P3 for salt precipitation and redissolve at 37 degrees C is necessary. | #Check Buffers P2 and P3 for salt precipitation and redissolve at 37 degrees C is necessary. | ||
*After preliminary steps have been done the miniprep is performed with the following steps: | *After preliminary steps have been done the miniprep is performed with the following steps: | ||
- | # | + | #Resuspend the pelleted bacterial cells in 250 microliters Buffer P1 and put in a centrifuge tube. |
#Add 250 microliters Buffer P2 and mix, invert 4-6 times. Solutions containing LyseBlue will turn blue. | #Add 250 microliters Buffer P2 and mix, invert 4-6 times. Solutions containing LyseBlue will turn blue. | ||
#Add 350 microliters N3 and IMMEDIATELY invert 4-6 times. Solutions with LyseBlue will turn colorless. | #Add 350 microliters N3 and IMMEDIATELY invert 4-6 times. Solutions with LyseBlue will turn colorless. |
Revision as of 21:24, 29 October 2008
ProtocolsMiniprep Prodedure
Restriction Digest
Gel Electrophoresis
Ligation Reaction Procedure
Transformation Procedure
Ethanol Precipitation
Gel Extraction
The contents of the tube should be yellow after incubation.
Calf CIP - Calf Intestinal Alkaline Phosphatase
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