Team:University of Lethbridge/Notebook/Project3October

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===October 7 2008===
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====Andrew, Nathan Puhl====
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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.
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[[Image:rpsA Gel Extraction.jpg|250 px]]
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===October 14 2008===
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====Andrew====
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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.
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No band present for the +1 TIR DNA, meaning gel extraction was not successful.
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===October 16 2008===
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====Andrew====
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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.
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Heat deactivated at 65 C after 1 hour incubation at 37 C.
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===October 21 2008===
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====Andrew====
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Transformation of +18 rpsA TIR DNA into pGEM T-easy to make enough DNA to be restricted/ligated into biobrick format. (psB1A2)
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50 uL og xgal added to LB + amp plates to screen for blue and white colonies.
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===October 22 2008===
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====Andrew====
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Transformation into pGEM T-easy successful, many white and some blue colonie on both plates.
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Picked 3 colonies from each plate to grow in 5mL LB + amp for a pDNA prep.
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===October 23 2008===
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====Andrew====
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pDNA prep of 6 culture tubes using a kit

Latest revision as of 02:23, 30 October 2008

Back to The University of Lethbridge Main Notebook


Contents

October 7 2008

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.


RpsA Gel Extraction.jpg


October 14 2008

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.


October 16 2008

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.


October 21 2008

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.


October 22 2008

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.


October 23 2008

Andrew

pDNA prep of 6 culture tubes using a kit

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