233
edits
SBB12Ntbk-JonathanKotker: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
→Jon Kotker, 2 March 2012
No edit summary |
|||
| Line 324: | Line 324: | ||
'''Part pBca9525-sbb1204''': ''Analytical Digests for Mapping'' | '''Part pBca9525-sbb1204''': ''Analytical Digests for Mapping'' | ||
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. | |||
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. | |||
'''Part pBca9525-sbb1223''': ''Analytical Digests for Mapping'' | '''Part pBca9525-sbb1223''': ''Analytical Digests for Mapping'' | ||
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. | |||
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. | |||
'''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]. | ||
