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Team:BYU Provo/Notebook/CholeraDetection/Springexp/Period1/Dailylog
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-KK Over the break we did little Lab work. Kelton was in Rexburg, and Clarice and I didn’t have the necessary primers to continue working. Today the primers came! We submitted primers for all the genes that have been cloned into pIG78 with pIG78 for sequencing. (I believe we included the primers for all the genes). We also have primers for working with CRO in the pBAD plasmid. Today our assignment was to make goals and set plans for what we hope to accomplish by the end of the term. Our plans are to be submitted by Friday. Our plans are spelled out in a table we’re printing, but they include understanding (via sequencing) what is ocurring in the plasmid by May 13th, correcting our system by May 29th, and demonstrating that we can induce Lambda into lysis through expression CRO. We will place CRO on the pBAD plasmid with the pBAD promoter (or on pLAT with a pBAD promoter), and the pBAD promoter is induced by arabinose. | -KK Over the break we did little Lab work. Kelton was in Rexburg, and Clarice and I didn’t have the necessary primers to continue working. Today the primers came! We submitted primers for all the genes that have been cloned into pIG78 with pIG78 for sequencing. (I believe we included the primers for all the genes). We also have primers for working with CRO in the pBAD plasmid. Today our assignment was to make goals and set plans for what we hope to accomplish by the end of the term. Our plans are to be submitted by Friday. Our plans are spelled out in a table we’re printing, but they include understanding (via sequencing) what is ocurring in the plasmid by May 13th, correcting our system by May 29th, and demonstrating that we can induce Lambda into lysis through expression CRO. We will place CRO on the pBAD plasmid with the pBAD promoter (or on pLAT with a pBAD promoter), and the pBAD promoter is induced by arabinose. | ||
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| - | - | + | -KP Today Kendall and I did PCR for the first time. Jordan and Clarice showed us how to do it. We did PCR on our E. Coli that has our lysogenic lambda encased within. We also froze down our lambda e. coli samples for future use. |
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| - | + | -KK Kelton and I worked with Jordan today to PCR amplify the CRO gene from π9907-infected E.Coli. We boiled the E.Coli to use as template and then followed the PCR protocol as outlined. Jordan actually was performed most of the protocol so that we could learn. Our control was E.Coli that had not been infected with lambda. | |
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Revision as of 22:10, 5 June 2013
| Cholera Detection May - June Notebook: May 1 - May 12 Daily Log
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5/1/13 -KK Over the break we did little Lab work. Kelton was in Rexburg, and Clarice and I didn’t have the necessary primers to continue working. Today the primers came! We submitted primers for all the genes that have been cloned into pIG78 with pIG78 for sequencing. (I believe we included the primers for all the genes). We also have primers for working with CRO in the pBAD plasmid. Today our assignment was to make goals and set plans for what we hope to accomplish by the end of the term. Our plans are to be submitted by Friday. Our plans are spelled out in a table we’re printing, but they include understanding (via sequencing) what is ocurring in the plasmid by May 13th, correcting our system by May 29th, and demonstrating that we can induce Lambda into lysis through expression CRO. We will place CRO on the pBAD plasmid with the pBAD promoter (or on pLAT with a pBAD promoter), and the pBAD promoter is induced by arabinose.
5/3/13 -KP Today Kendall and I did PCR for the first time. Jordan and Clarice showed us how to do it. We did PCR on our E. Coli that has our lysogenic lambda encased within. We also froze down our lambda e. coli samples for future use. -KK Kelton and I worked with Jordan today to PCR amplify the CRO gene from π9907-infected E.Coli. We boiled the E.Coli to use as template and then followed the PCR protocol as outlined. Jordan actually was performed most of the protocol so that we could learn. Our control was E.Coli that had not been infected with lambda.
5/4/13 - Performed dilution series using stock 5.3 (-1 through -11) - Started two 5mL overnights of BL21
5/5/13 - Spot test using stock 5.3 and its dilution series (from 5.4) - Started liquid culture for purification team (at around noon)
- Started 5.5 amplification from a plaque test
5/6/13 - Continued 5.3 T7 phage amplification/purification - Performed liquid culture phage concentration test
5/7/13 - Started two 5mL of E coli BL21 overnight - Designed procedure for applying mutagen and selecting for T7
5/8/13 - Went over procedure for applying mutagen and PCR with Dr. Grose - Performed spot tests under 5.6 T7+ Liquid Culture Phage Concentration Test - Started two 5mL BL21 overnights - Learned how to create LB plates
5/9/13 - Practiced with ×6 and ×8 top agar - Started 5.9 T7+ Liquid Culture Phage Concentration Test #2 - Discussed plans and schedule for next week.
5/10/13 - Worked on Progress Report
5/12/13 - Started two 5mL overnight at around 6pm
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