Team:University of Ottawa/31 July 2008

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 Home  Welcome Announcements</li> </ul> The Team</a> <ul> Who We Are</a> <ul> Advisors</a></li> Undergrads</a></li> </ul> </li> What We've Done</a></li> Where We're From</a></li> Contact Us</a></li> </ul> </li> The Project</a> <ul> <li><a href="http://2008.igem.org/Team:University_of_Ottawa/Project#Project_Overview">Overview</a></li> <li><a href="http://2008.igem.org/Team:University_of_Ottawa/Project#Pulsate_Gene_Expression">Expression</a></li> <li><a href="http://2008.igem.org/Team:University_of_Ottawa/Project#Cell-to-Cell_Communication">Communication</a></li> <li><a href="http://2008.igem.org/Team:University_of_Ottawa/Project#Oscillatory_Dynamics">Oscillation</a></li> <li><a href="http://2008.igem.org/Team:University_of_Ottawa/Project#Applications">Application</a></li> <li><a href="http://2008.igem.org/Team:University_of_Ottawa/Project#Journal_Club">Journal Club</a></li>

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Chris
Microwave Digestion Experiment
 * <li> Following protocol outlined online, tested a microwave protocol to hasten digestion period.
 * <li> Digested DQ232597 with HindIII. Ran five samples: 5 seconds in micro, 10 seconds, 15 seconds, 10 seconds in and 2 minutes out (four times), water control, water control 10 seconds in microwave and 2 minutes out, 1 hr traditional digest.
 * <li> Denatured HindIII at 80 C for 20 minutes, then ran products on a gel.
 * <li> Results promising; however, more experimentation needed to confirm validity of this protocol

Matt
Ligation
 * <li> Another ligation of PTP2 insert with pSSA42 vector was performed with 3:1 ratio, 6:1 ratio, Vector with ligase, vector without ligase and H2O control.
 * <li> 2 hr ligation was unsucessful, overnight ligation was usccessful for both 3:1 and 6:1.

Transformation
 * <li> Transformed ligation product into competent cells - 3 different plates -3:1, 6:1 and control.

Dan
Gel Confirmation
 * <li> One gel was run for the confirmation of the ligation of constructs 0A 0B and T, this gel showed that the ligation was successful

Gel Extraction
 * <li> The successful gel indicated that I could go forward and run all the samples on a gel for gel extraction (ran over 6 wells) and the correct bands were extracted.

Digestion
 * <li> 2 samples were created (one for gel confirmation and one for transformation), both were digested with ClaI

PCR cleanup
 * <li>PCR cleanup of the digestion was performed in order to prevent salts from inhibiting gel electrophoresis.

Ligation
 * <li> Ligation was run overnight in the thermocycler of pSSA140B and p1T(0A0B)