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Team:Calgary/2 August 2010
From 2010.igem.org
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Set up a restriction digest of pRFP without PCR purification and left overnight. This was cut with EcoRI and SpeI. Since the sample had salts from PCR, concentration was not measured therefore a estimation of around 5.5uL to make the total concentration of the sample 10 uL. A PCR purification was made using older sample however even combining all the wells making the DNA volume in the tube 100uL, the concentrations were in their negatives. PCR purification will not be pursued further. Attempts will be made to plasmid switch PCR product without purification. | Set up a restriction digest of pRFP without PCR purification and left overnight. This was cut with EcoRI and SpeI. Since the sample had salts from PCR, concentration was not measured therefore a estimation of around 5.5uL to make the total concentration of the sample 10 uL. A PCR purification was made using older sample however even combining all the wells making the DNA volume in the tube 100uL, the concentrations were in their negatives. PCR purification will not be pursued further. Attempts will be made to plasmid switch PCR product without purification. | ||
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| + | <u>Dev</u> | ||
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| + | Obtained no colonies from yesterday's plasmid switch. Decided to re-ligate the digested insert and vector from yesterday, transform and plate. | ||
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Revision as of 16:56, 5 August 2010
Monday August 2, 2010
Heritage Day: A Day off!!
Jeremy
Set up a restriction digest of pRFP without PCR purification and left overnight. This was cut with EcoRI and SpeI. Since the sample had salts from PCR, concentration was not measured therefore a estimation of around 5.5uL to make the total concentration of the sample 10 uL. A PCR purification was made using older sample however even combining all the wells making the DNA volume in the tube 100uL, the concentrations were in their negatives. PCR purification will not be pursued further. Attempts will be made to plasmid switch PCR product without purification.
Dev
Obtained no colonies from yesterday's plasmid switch. Decided to re-ligate the digested insert and vector from yesterday, transform and plate.
No notebook page exists for this date. Sorry!
