Team:University of Lethbridge/Notebook/Project3October

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Andrew, Nathan Puhl
Ran a gel to confirm presence of DNA from PCR on September 30 2008. DNA present in all wells, so a 2% agaraose gel was prepared for a gel extraction of DNA. 40uL of DNA was loaded and gel extracted using a kit.



Andrew
Ran 1uL of gel extraction products on a gel to determine presence/quantity of DNA.

Band close to ~150 bp for the +18 rpsA DNA, which is approximately the correct size. No band present for the +1 TIR DNA, meaning gel extraction was not successful.

Andrew
Ligation of +18 rpsA DNA into pGEM T-easy beacuse primer design requires DNA to be in a plasmid for endonuclease activity of restriction enzymes.

Heat deactivated at 65 C after 1 hour incubation at 37 C.

Andrew
Transformation of +18 rpsA TIR DNA into pGEM T-easy to make enough DNA to be restricted/ligated into biobrick format. (psB1A2)

50 uL og xgal added to LB + amp plates to screen for blue and white colonies.

Andrew
Transformation into pGEM T-easy successful, many white and some blue colonie on both plates.

Picked 3 colonies from each plate to grow in 5mL LB + amp for a pDNA prep.

Andrew
pDNA prep of 6 culture tubes using a kit