5. If a sample of double-stranded DNA gave the following results, what is the co

Question

5. If a sample of double-stranded DNA gave the following results, what is the concentration (ng/ul) of the sample? How much DNA would you add to your PCR reaction to achieve the recommended 100 ng of DNA template? Show your work. (2 points) 50g/mL 1.456 72,Sm/mL = A260 = 1.456 A230 = 0.975 A2801.017 · · Using the concentration (ng/W) that you calculated in #5, what is the total amount of dsDNA in micrograms (ug) that you extracted if your total sample's volume was 250 ul? Show your work. (1 point)

Explanation / Answer

5) The absorption value of 1 at A260 corresponds to 50 ug/ml

So, the value of 1.456 at A260 corresponds to,

= 50 x 1.456

= 72.8 ug/ml

The amount of DNA added to PCR reaction to achieve concentration of 100 ng is,

= (72.8 ug x 1000 ng x 1 ml)/(1 ml x 1 ug x 1000 ul)

= 72.8 ng/ul

6) If the volume is 250 ul, the amount of dsDNA is,

= 72.8 x 250 ng

= 18200 ng

= 18.2 ug

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