Team:University of Ottawa/7 August 2008

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Latest revision as of 15:24, 12 August 2008

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Today in the Lab

Chris

Digestion and Ligation of OA, OB and T Amp Product

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°C for 15 minutes, followed by 20 minutes at 80°C to denature.

Spiked each tube with ligase and ATP then incubated at room temperature for around two hours.

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.

Excised desired band and performed gel extraction. Stored at -20°C.

Matt

Absorbance was measured from the ten colonies miniprepped by Chris. I then digested each colony with PST1 to confirm.

All colonies were confirmed and successful.

Used SLN1 primers to amplify the PTP2/GAL10 casette overnight from two colonies chosen from the miniprep.

Dan

p1T miniprep confirmation

p1T was digested with HindIII for confirmation, I did not get the correct product :(

dehydrogenase construct

pSSB37 and pDR197(pure1) were both digested with SphI and HindIII

Master plates of pSSB37 and pDR197(pure1) were streaked

@@ Line 84: / Line 84: @@
 :<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> I used SLN1 primers to amplify the PTP2/GAL10 casette overnight from two colonies chosen from the miniprep.
+:<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