Team:BYU Provo/Notebook/CholeraDetection/Springexp/Period3/Dailylog

From 2013.igem.org

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- Discussed plans for the selection process of [[Team:BYU Provo/Notebook/SmallPhage/Springexp/Period2/Exp/5.20 Mutagen Concentration Experiment|5.20 Mutagen Concentration Experiment]] and calculated the needed volume of agar, overnight, and phage
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Regarding our PCR attempt yesterday, we did see a thick fat band ... at the bottom of the gel! Once again our PCR failed. This has led us to propose several reasons why we can't PCR amplify CRO: 1) the Lambda prophage isn't actually in TT 9901 or TT9907, 2)
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- Made 500mL x8 agar
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- Performed dilution series to generate enough 200ug -4 phage stock for selection.
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Revision as of 19:03, 7 June 2013


Cholera Detection May - June Notebook: May 27 - June 9 Daily Log



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

5/28/13

- Started three 8mL E coli BL21 liquid culture at around 4pm.


5/29/13

- Continued 5.20 Mutagen Concentration Experiment

- Prepared sample for sequencing. This is done as part of 5.20 T7 Minor Capsid Protein PCR

- Dr. Studier responded to our email today! We also received the E coli stains containing plasmids with clone T7 genes. WE CAN START SITE DIRECTED MUTAGENESIS SOON!


5/30/13

- Plates from yesterday are taken out of incubation at around 4:00pm


5/31/13

- Discussed results for 5.20 Mutagen Concentration Experiment -> will need to try this one more time with minor adjustment to experimental design: plate -4 dilution using x6 and x8 top agar.

- Discussed plans for next week.

- Made new LB and x6 top agar.


6/2/13

- Made about 20ml of BL21 overnight


6/3/13

- Made new LB plates

- Plated -4 titer on x6 and x8 plates for 5.20 Mutagen Concentration Experiment


6/5/13

Regarding our PCR attempt yesterday, we did see a thick fat band ... at the bottom of the gel! Once again our PCR failed. This has led us to propose several reasons why we can't PCR amplify CRO: 1) the Lambda prophage isn't actually in TT 9901 or TT9907, 2)


5/20/13

- Performed T7 Mutagen Concentration Test

5.20 Mutagen Concentration Experiment

- Performed T7 Minor Capsid Protein PCR

5.20 T7 Minor Capsid Protein PCR


5/21/13

- Started two 5mL E coli BL21 overnight at around 7:00pm


5/22/13

- Performed spot test for 5.20 Mutagen Concentration Experiment

- Ran agarose gel to confirm PCR product

5.20 T7 Minor Capsid Protein PCR


5/23/13

- Started two 25mL E coli BL21 liquid culture over night at around 6:00pm


5/24/13

- Proceeded with 5.20 Mutagen Concentration Experiment by performing preliminary selection using x8 top agar


5/25/13

- Took pictures in preparation for Progress Report


5/31/13

- Worked on transferring our notebook over to the iGEM wiki.