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You performed a miniprep and put the DNA into Tube 1, containing a total volume

ID: 149977 • Letter: Y

Question

You performed a miniprep and put the DNA into Tube 1, containing a total volume of 450l. You then took 10l of the DNA from Tube 1 and added it to 70l of water in Tube 2. You then measured the absorbance of Tube 2 and found A260 = 0.298 and A280 = 0.154.

A) What is the concentration of your miniprep DNA in Tube 1 (in g/mL)?

B) What is the concentration of your miniprep DNA in Tube 2 (in g/mL)?

C) What is your total DNA yield?

D) What can you specifically say about the purity of your plasmid preparation?

Explanation / Answer

dsDNA concentration = 50 g/mL × OD260 × dilution factor.

Ans B

as you took 10ul dna and diluted with 70ul water then dilution factor is 8

on applying in the formula we get.

dsDNA conc= 50*0.298*8= 119.2

ANS a

now simply use the formula C1v1=c2v2

c1*10=119.2*80

c1=953.6

0.953mg/ml

ans c total conc yield = 0.953mg/ml

D- The plasmid is of good quality. Because a260/a280 ratio should have in between 1.7-2.0

0.298/0.154= 1.93

Very Good qualtiy

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