Team:BYU Provo/Notebook/CholeraDetection/Summerexp/Period2/Dailylog

Cholera Detection July-August Notebook: July 15 - July 28 Daily Log

7/15/2013

Tasks: Do safety paperwork. Research how to induce lytic phage in lamdba.

Set up of phage test: Made overnight cultures of the three strains from the Roth Lab and the two strains that have pLAT+Cro. Grown in LB-Am. Plate 100ul onto LB or LB-Am. Spot Hydrogen Peroxide onto the middle of the plate. Put at 37. Check for phage lysis the next day.

KP,KK,CH

7/16/13
Copies of RFP in the iGEM data base to use as template- 2012 Plate 1>18F, 2012 Plate 1>220, 2011 Plate SP4001>11H.

Redid PCR from 7/12 using all the templates above and also pSB1C3 (polymerase phusion). The reaction using pSB1C3 as template was the only one that worked. Cleaned it. Resulted in promoter Qrr4 reading into RFP.

Made a mini prep of pIG10 (AmR).

Set up 50ul digests of Qrr4/RFP and pIG12 with NEB4, XhoI, and EcoRI-HF. Ran on a low melt gel. The band with Qrr4/RFP was dim and the band with pIG12 was bright.

KK, KP, CH

7/19/13

Set up ligations of pIG12 with Qrr4/RFP (digested 7/18). Also set up a vector only control. Transformed into dH5alpha. There were no colonies on our vector only control. Collected 10 or so colonies from the vector plus insert ligation.

KK, KP, CH

7/22/13
Set up a PCR check (taq polymerase) for our pIG10 Qrr4/RFP colonies. Colonies E,F, and G gave the best bands for inserts.

KK, KP, CH

7/24/13

Set up overnights yesterday of Colonies E,F and G. Today mini preped them. Submitted for sequencing using vector primers IG11 (forward) and IG12 (reverse). 2ul sample with 1ul primer. All clones had perfect sequence.

KK, KP, CH