Team:University of Ottawa/2 July 2008

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(New page: __TOC__ ==Today in the Lab== '''Matt''' :'''Ligation''' ::<li> I am using 2:1, 3:1, 4:1 molar ratio of insert to vector being used for ligation of PTP2 to pSSA42. ::<li> A gel confirmation...)
 
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__TOC__
__TOC__
==Today in the Lab==
==Today in the Lab==
'''Matt'''
'''Matt'''
:'''Ligation'''
:'''Ligation'''
-
::<li> I am using 2:1, 3:1, 4:1 molar ratio of insert to vector being used for ligation of PTP2 to pSSA42.
+
::<li> I am using three different samples of 2:1, 3:1, 4:1 molar ratio of insert to vector being used for ligation of PTP2 to pSSA42.  
::<li> A gel confirmation was run however nothing appeared on the gel due to a very low DNA concentration of the PTP2.
::<li> A gel confirmation was run however nothing appeared on the gel due to a very low DNA concentration of the PTP2.
::<li> We decided it was better just to try and integrate into competent cells and incubate overnight for tomorrow morning.
::<li> We decided it was better just to try and integrate into competent cells and incubate overnight for tomorrow morning.
:'''Transformation'''
:'''Transformation'''
-
::<li> The competent cells were transformed with the ligation product and left overnight.The receptor component Atcre was also integrated into competent cells.
+
::<li> The competent cells were transformed with the ligation product of all three samples and left overnight.The receptor component Atcre was also integrated into competent cells.
:'''Dehydrogenase component'''
:'''Dehydrogenase component'''
::<li> The dehydrogenase was received in the mail, I streaked the Ecoli containing the component to grow overnight in the incubator. Tomorrow we will inoculate for mini prep the following day.
::<li> The dehydrogenase was received in the mail, I streaked the Ecoli containing the component to grow overnight in the incubator. Tomorrow we will inoculate for mini prep the following day.
 +
'''Dan'''
 +
:'''PCR of pSSRE constucts'''
 +
::<li> Turned out to be a disaster. Had nonspecific binding and >4 bands. We would be better of going back to the original gel extraction products.
 +
:'''Digestion'''
 +
::<li> Now I am using the original gel extraction products (1,2,3,4) and

Latest revision as of 23:41, 28 October 2008

Untitled Document

 

 

Contents

Today in the Lab

Matt

Ligation
  • I am using three different samples of 2:1, 3:1, 4:1 molar ratio of insert to vector being used for ligation of PTP2 to pSSA42.
  • A gel confirmation was run however nothing appeared on the gel due to a very low DNA concentration of the PTP2.
  • We decided it was better just to try and integrate into competent cells and incubate overnight for tomorrow morning.
    • Transformation
  • The competent cells were transformed with the ligation product of all three samples and left overnight.The receptor component Atcre was also integrated into competent cells.
    • Dehydrogenase component
  • The dehydrogenase was received in the mail, I streaked the Ecoli containing the component to grow overnight in the incubator. Tomorrow we will inoculate for mini prep the following day.

    • Dan

    PCR of pSSRE constucts
  • Turned out to be a disaster. Had nonspecific binding and >4 bands. We would be better of going back to the original gel extraction products.
    • Digestion
  • Now I am using the original gel extraction products (1,2,3,4) and
    • Retrieved from "http://2008.igem.org/Team:University_of_Ottawa/2_July_2008"