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Team:British Columbia/Notebook/27 July 2011
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| - | *The previous colonies of E. coli that should have had the 1,8-cineole gene on pSB1C3 were shown to have different sized plasmids after overnight cultures and minipreps. Of the 5 colonies, 4 had plasmids | + | *The previous colonies of E. coli that should have had the 1,8-cineole gene on pSB1C3 were shown to have different sized plasmids after overnight cultures and minipreps. Of the 5 colonies, 4 had plasmids a little over 2 kb in size, and one had a plasmid around 5 kb in size. The 1,8-cineole gene is 1.7 kb and the pSB1C3 plasmid that is left after the restriction digest is 2 kb, so the product should have been around 3.7 kb. A restriction digest with ECORI and PSTI showed only one band for the 5 kb product, and the 2 kb product showed bands at about 1.7 kb and 700 bp. All of these were grown on chlor plates and chlor LB broth for the overnight culture. A gel of just the pSB1C3 showed a product well below the 500 bp mark on 1 kb ladder. |
*This was rather disconcerting, so Jacob decided to run another biobrick PCR and try the ligation again. | *This was rather disconcerting, so Jacob decided to run another biobrick PCR and try the ligation again. | ||
Latest revision as of 17:52, 2 August 2011
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July 27 2011
1,8-cineole
- The previous colonies of E. coli that should have had the 1,8-cineole gene on pSB1C3 were shown to have different sized plasmids after overnight cultures and minipreps. Of the 5 colonies, 4 had plasmids a little over 2 kb in size, and one had a plasmid around 5 kb in size. The 1,8-cineole gene is 1.7 kb and the pSB1C3 plasmid that is left after the restriction digest is 2 kb, so the product should have been around 3.7 kb. A restriction digest with ECORI and PSTI showed only one band for the 5 kb product, and the 2 kb product showed bands at about 1.7 kb and 700 bp. All of these were grown on chlor plates and chlor LB broth for the overnight culture. A gel of just the pSB1C3 showed a product well below the 500 bp mark on 1 kb ladder.
- This was rather disconcerting, so Jacob decided to run another biobrick PCR and try the ligation again.
