Team:University of Ottawa/7 August 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>

</ul> </li> <li><a href="http://2008.igem.org/Team:University_of_Ottawa/Parts">BioBricks</a></li> <li><a href="http://2008.igem.org/Team:University_of_Ottawa/Modeling">Modeling</a></li> <li><a class="MenuBarItemSubmenu" href="http://2008.igem.org/Team:University_of_Ottawa/Wet_Lab">Wet Lab</a> <ul> <li><a href="http://2008.igem.org/Team:University_of_Ottawa/Lab_Protocols">Lab Protocols</a></li> <li><a href="http://2008.igem.org/Team:University_of_Ottawa/WetWare">WetWare</a></li> </ul> <li><a href="http://2008.igem.org/Team:University_of_Ottawa/Notebook">Notebook</a></li> <li><a class="MenuBarItemSubmenu" href="http://2008.igem.org/Team:University_of_Ottawa/Sponsors">Sponsors</a> <ul> <li><a href="http://2008.igem.org/Team:University_of_Ottawa/Sponsors#Acedemic_Sponsors">Academic</a></li> <li><a href="http://2008.igem.org/Team:University_of_Ottawa/Sponsors#Research_Sponsors">Research</a></li> <li><a href="http://2008.igem.org/Team:University_of_Ottawa/Sponsors#Corporate_Sponsors">Corporate</a></li> </ul> </li>

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Chris
Digestion and Ligation of OA, OB and T Amp Product
 * <li> Digested all three according to previous digestion protocol (see August 5 post). Ran 7 samples--six ligations and one digestion products only. Incubated at 37&deg;C for 15 minutes, followed by 20 minutes at 80&deg;C to denature.
 * <li> Spiked each tube with ligase and ATP then incubated at room temperature for around two hours.
 * <li> Ran products on a 0.6% gel for 1.5 hours at 80V. Stained the gel in an Ethidium Bromide and TAE Buffer solution for 20 minutes.
 * <li> Excised desired band and performed gel extraction. Stored at -20&deg;C.

Matt

 * <li> Absorbance was measured from the ten colonies miniprepped by Chris. I then digested each colony with PST1 to confirm.
 * <li> All colonies were confirmed and successful.
 * <li> Used SLN1 primers to amplify the PTP2/GAL10 casette overnight from two colonies chosen from the miniprep.

Dan
p1T miniprep confirmation
 * <li> p1T was digested with HindIII for confirmation, I did not get the correct product :(

dehydrogenase construct
 * <li> pSSB37 and pDR197(pure1) were both digested with SphI and HindIII
 * <li> Master plates of pSSB37 and pDR197(pure1) were streaked