3. Use the following agarose gel and information on page 44 of your lab manual t

Question

3. Use the following agarose gel and information on page 44 of your lab manual to answer the questions below Lane 1+12: 1 kb DNA ladder Lanes 2-9: PCR products (5 L each) Lane 10: 16S negative control Lane 11: rpoB negative control Lane 2 contains the (circle): 16s/ rpoB PCR fragment. The molecules in this band are long. The mass of DNA in this band is into lane 2 was bp a. ng and the DNA concentration of the PCR that was loaded ng/|. To set up a DNA sequencing reaction with 75 ng DNA, you need to add ul ofyour PCR, 1 1 of primer and ddH2O to a total volume of 13 Lane 3 contains the (circle): 16s / rpoB PCR fragment. The molecules in this band are long. The mass of DNA in this band is bp b. ng and the DNA concentration of the PCR that was loaded o lane 3 was ng/. To set up a DNA sequencing reaction with 75 ng DNA, you need to add 1 of your PCR, 1 1 of primer and ddH2O to a total volume of 13 .

Explanation / Answer

a. The molecule in this band are 900 bp in length. The mass of the DNA band is 60.48 ng and the concenration of the PCR product that was loaded into lane 2 was 12ng/l. To set up a DNA sequencing reaction with 75ng DNA,you need to add 6.25l of your PCR, 1l of primer and ddH2O to a total volume of 13l.

b. the molecule in this band are 500 bp in length. The mass of the DNA band is 67.2ng and the concentration of the PCR product that was loaded into lane 3 was 13.4ng/ l. To set up a DNA sequencing reaction with 75ng, you need to add 5.5l. of your PCR, 1l. of primer and ddh20 to a total volume of 13l.

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