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SBB12Ntbk-JonathanKotker: Difference between revisions
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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. This was used on the colonies that I picked [http://openwetware.org/wiki/SBB12Ntbk-JonathanKotker#Jon_Kotker.2C_1_March_2012 yesterday]. | 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. This was used on the colonies that I picked [http://openwetware.org/wiki/SBB12Ntbk-JonathanKotker#Jon_Kotker.2C_1_March_2012 yesterday]. | ||
==[[User:Jonathan_Kotker|Jon Kotker]], 6 March 2012== | |||
'''Part pBca9525-sbb1223''': ''Analytical Digests for Mapping'' | |||
The procedure is described [http://openwetware.org/wiki/Template:SBB-Protocols_Enz6 here] and is used to guess if the purified product is indeed the DNA sequence we need. We pick particular restriction enzymes such that the resulting fragments are of significantly different sizes, which should thus show up as two distinct bands on the analytical gel. If the analytical gel shows bands at the approximately proper locations, then we can send the purified product for digestion. | |||
Based on the construction file for this part, I chose the restriction enzymes EcoRI and BamHI, which should result in parts of length 1400bp and 2472bp. The procedure is modified accordingly: | |||
Recipe: | |||
* 4uL ddH2O | |||
* 4uL 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 10:48am. (I took it out at 11:50am.) | |||
