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SBB12Ntbk-JonathanKotker: Difference between revisions
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# Drop it in the test tube. | # Drop it in the test tube. | ||
# Incubate at 37 overnight. | # Incubate at 37 overnight. | ||
'''Part pBca9525-sbb1204''': ''Miniprep Purification of DNA'' | |||
The procedure is described [http://openwetware.org/wiki/Template:SBB-Protocols_Micro3 here] and is used to extract and purify the DNA retrieved from the cells grown on [http://openwetware.org/wiki/SBB12Ntbk-JonathanKotker#Jon_Kotker.2C_8_March_2012 March 8]. | |||
'''Part pBca9525-sbb1204''': ''Analytical Digests for Mapping'' | |||
Based on the construction file for this part, I chose the restriction enzymes EcoRI and XhoI, which should result in parts of length 2634bp and 986bp. The procedure is modified accordingly: | |||
Recipe: | |||
* 7uL ddH2O | |||
* 1uL Miniprepped plasmid | |||
* 1uL 10x NEB Buffer 2 | |||
* 0.5uL EcoRI | |||
* 0.5uL BamHI | |||
Procedure: | |||
# Incubate at 37 degrees on the thermocycler for 30 minutes after 1:08pm. | |||
The result is shown [http://openwetware.org/wiki/Image:2012_03_20_gel3_ssb2012spring.jpg here] and replicated below for completeness: | |||
[[Image:NEB_2-log_ladder.gif]][[Image:2012_03_20_gel3_ssb2012spring.jpg|500px]] | |||
My two samples (from colonies 3 and 4) were added to the seventh and eighth lane from the left. | |||
