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SBB12Ntbk-JonathanKotker: Difference between revisions
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→Jon Kotker, 2 March 2012
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The procedure is described [http://openwetware.org/wiki/Template:SBB-Protocols_Micro3 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. Unfortunately, I only got as far as digesting some of the purified product, since there were no gels available. | The procedure is described [http://openwetware.org/wiki/Template:SBB-Protocols_Micro3 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. Unfortunately, I only got as far as digesting some of the purified product, since there were no gels available. | ||
Based on the construction file for this part, I chose the restriction enzymes EcoRI and XhoI, which should result in parts of length bp and bp. | Based on the construction file for this part, I chose the restriction enzymes EcoRI and XhoI, which should result in parts of length bp and bp. The incubation period was 30 minutes after 1:08pm. | ||
'''Part pBca9525-sbb1223''': ''Analytical Digests for Mapping'' | '''Part pBca9525-sbb1223''': ''Analytical Digests for Mapping'' | ||
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The procedure is described [http://openwetware.org/wiki/Template:SBB-Protocols_Micro3 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. Unfortunately, I only got as far as digesting some of the purified product, since there were no gels available. | The procedure is described [http://openwetware.org/wiki/Template:SBB-Protocols_Micro3 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. Unfortunately, I only got as far as digesting some of the purified product, since there were no gels available. | ||
Based on the construction file for this part, I chose the restriction enzymes EcoRI and BamHI, which should result in parts of length bp and bp. | Based on the construction file for this part, I chose the restriction enzymes EcoRI and BamHI, which should result in parts of length bp and bp. The incubation period was 30 minutes after 1:08pm. | ||
'''Part pBca9525-sbb1223''': ''Miniprep Purification of DNA'' | '''Part pBca9525-sbb1223''': ''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. 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]. | ||
