If a sample of double-stranded DNA was diluted 20X, and gave an A26o reading of

Question

If a sample of double-stranded DNA was diluted 20X, and gave an A26o reading of 0.650D on a spectrophotometer, using the formula above, what is the concentration of dsDNA in the original sample? Show your work (2 points): ds DNA concentration-50 ?g/ml x 0D24ox dilution tactor 20x :ds DNA concentraion 50gn Now that you calculated the dsDNA concentration (ug/ml) in your sample, you can determine the total dsDNA yield (ug) in your extraction by multiplying the two values together. 6. If you had eluted the DNA sample used in question 5 in 300 ul of elution buffer calculate the total amount of DNA (18) you extracted from your sample. Show your work and include the correct units in your final answer. (2 poihts)

Explanation / Answer

5. the concentration of DNA can be measured by absorbance at 260nm since the DNA shows maximum absorption at 260nm. I unit of absorbance will have 50 µg of ds DNA.

So the formula can be given as

Concentration (µg/ml) = A260 reading × dilution factor × 50µg/ml

= 0.65 * 20 * 50=650 µg/ml

6. elution buffer is 300 microliters so this is 3 ml

DNA yield (µg) = DNA concentration × total sample volume (ml)

= 650 * 3

=1950 µg

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