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

From 2013.igem.org

(Difference between revisions)

Revision as of 19:14, 7 June 2013

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

5/28/13

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

5/29/13

-KP KK Today we plated TT9907 and TT9901. We plated them on LB arabinose/amp/x-gal plates. We plated different concentrations of TT9907 and TT9901 on each of the plates. We added 50, 100, and 300 microliters onto each plate. We will check the plates tomorrow hopefully to find plagues showing that arabinose triggered lysis of our lambda. We unfortunately didn't get any growth of our TT23821, so we streaked another plate from our original stab sample and we will try to re-electroporate TT23821 on Friday.

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, or 2) our primers aren’t functioning to amplify CRO. We streaked a few plates of TT9901 and TT9907 to subject to UV light. If we see plaques, then we will know Lambda has been induced. Meanwhile Dr. Grose is going to check that our primers are the correct ones.

@@ Line 38: / Line 38: @@
 <font size="4"> '''5/29/13''' </font>
-- Continued [[Team:BYU Provo/Notebook/SmallPhage/Springexp/Period2/Exp/5.20 Mutagen Concentration Experiment|5.20 Mutagen Concentration Experiment]]
+-KP KK Today we plated TT9907 and TT9901. We plated them on LB arabinose/amp/x-gal plates. We plated different concentrations of TT9907 and TT9901 on each of the plates. We added 50, 100, and 300 microliters onto each plate. We will check the plates tomorrow hopefully to find plagues showing that arabinose triggered lysis of our lambda. We unfortunately didn't get any growth of our TT23821, so we streaked another plate from our original stab sample and we will try to re-electroporate TT23821 on Friday.
-- Prepared sample for sequencing. This is done as part of [[Team:BYU Provo/Notebook/SmallPhage/Springexp/Period2/Exp/5.20 T7 Minor Capsid Protein PCR|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!
 <br>