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Determine the stock concentration of template dna

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Reference no: EM132598815

You have a 50 µl of pEGFP template DNA from which you want to amplify the GFP ORF. Previously an aliquot of this DNA was diluted 1/100 in H2O and the absorbance at 260nm was measured using a nanodrop spectrophotometer. The absorbance (minus blank) was 0.34. (Note - an absorbance of 1 at 260nm corresponds to a concentration of 50 μg/ml for double-stranded DNA).

You need to add 25 ng of pEGFP template DNA in total to your reaction. Determine the stock concentration of your template DNA and come up with a protocol for diluting it to an appropriate concentration to allow you to add 25 ng to your PCR reaction - the volume of pEGFP should be no greater than 5 µl.

Reference no: EM132598815

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