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.
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Revision as of 22:08, 5 June 2013


Cholera Detection May - June Notebook: May 1 - May 12 Daily Log



Overview
March-April
May-June
July-August
September-October

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.


5/2/13

- Designed primers for amplifying and sequencing phage capsid protein

- Wrote emails inquiring mutagen (NG) and plasmid for in vitro assembly


5/3/13

- Performed agar test, focusing primarily on ×8

5.3 T7 phage selection method test

- Processed phage amplification

5.3 T7 phage amplification/purification


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)

5.3 T7 phage amplification/purification

- Started liquid culture for purification team (at around noon)

1mL of BL21 overnight + 4mL of LB + 100μL -2 phage (received stock)

- Started 5.5 amplification from a plaque test

5.5 Amplification from a plaque test


5/6/13

- Continued 5.3 T7 phage amplification/purification

- Performed liquid culture phage concentration test

5.6 T7+ 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

5.9 T7 Selection Method Test #2

- 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