Reference no: EM132800274
To check whether your revised PCR was successful, you load and run an agarose gel with molecular weight standards (lane MW) and your PCR reaction. Two other summer students in the lab ask if they can run samples from their experiments on your gel, and you allow them to do so. However, in your haste, you failed to record who loaded in which lane. The distance of migration for all bands are indicated.
Size (bp) Lane Distance Migrated (cm)
10000 MW 1.1
8000 MW 1.2
6000 MW 1.6
5000 MW 1.7
4000 MW 2.2
3000 MW. 2.6
2000 MW 3.6
1500 MW 4.5
1000 MW 5.9
500 MW 9.0
Unknown 1 7.9
Unknown 2 9.2
Unknown 3 6.6
a. Construct a standard curve in Excel that relates the distance of migration in centimeters to the log of DNA fragment length in basepairs (bp). Note that the log of fragment length is linearly related to the distance migrated. Please treat the distance migrated as the independent variable, do not round any values, and do not exclude pertinent data in the construction of your standard curve. Only provide the equation of the linear regression line for grading, in the standard form y=mx+b. Submissions will be penalized for extraneous information.
b. Given this standard curve, what are the sizes (in bp) of the DNA fragments in each of the three lanes? Round to the nearest whole number if necessary. Submissions will be penalized for extraneous information.
c. Out of these three predicted DNA fragment sizes, are there any values you are not confident about? If so, why? Answer in two grammatically correct sentences at most.
d. Which lane do you think contains your PCR reaction? Why? Answer in one grammatically correct sentence at most.