Team:University of Ottawa/21 July 2008

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__TOC__
__TOC__
==Today in the lab==
==Today in the lab==

Latest revision as of 18:30, 30 July 2008

Untitled Document

 

 


Contents

Today in the lab

Dan

Gel of sample 2T
  • For some odd reason all of the DNA stayed in the well... Could be some sort of salt contamination from NEB buffer 1, or a very large autoligation product. I will test the former by doing PCR cleanup and switching to ligation buffer and then run the gel again.
    • Digestion of 1A 0B and both S and D
  • The above DNA fragments were digested with PstI and SphI in Neb buffer 1, enzymes were denatured after an hour
  • PCR cleanup was performed on the digested products, eluted with 50 uL water.
    • Ligation
  • Ligation of the DNA obtained from PCR cleanup was run overnight at 16 C

    • Matt

    Miniprep
  • I performed a miniprep of the inoculated PTP2 plasmids and yielded good concentrations for all samples.
  • I then proceeded to digest each sample with NcoI and run them on a gel for confirmation.
  • It appeared as though NcoI only cleaved at one site on the plasmid - linearizing the plasmid. I am going to try to attempt another digestion with EcoRV/EagI which should give two definitive bands - if successful then I will PCR amplify with SLN1 primers for integration into yeast.

    • Chris

    Transformation of Competent Cells
  • Using ligated AtCRE that Dan prepared 22 July 2008, performed a transformation of competent cells according to protocol
  • Performed transformation on a single sample only, as success was not expected. Used LB plates with ampicillin. Incubated against a control plate with no e. coli.
  • Incubated overnight.
    • Retrieved from "http://2008.igem.org/Team:University_of_Ottawa/21_July_2008"