In a lab experiment, mitochondrial DNA from the members of the class was taken a
ID: 216325 • Letter: I
Question
In a lab experiment, mitochondrial DNA from the members of the class was taken and purified. The contol reegion of the mitochondrial genome was then amplified using PCR for each member of the class. It was amplified using the forward and reverse primers for two different PCR reactions; the primers used was the same for each individual, therefore everyone had the same forward primer and everyone had the same reverse primer. In the first PCR reaction only a portion (about 440 bp) of the control region was amplified. In the second PCR reaction the entire control region (about 1100 bp) was amplified. The PCR product was then analyzed using agarose gel electrophoresis.
How do I know if i amplified the correct region? Why are some band sizes the same and why re some different? Can i say that individuals are different or similiar based on these results?
This is a picture of the gel
Kim Kim Alexis Alexis Anna Anna Bitaax Britany DNA A B er ?.ABd Ladd DNA Wan Wan Ladder A BExplanation / Answer
for the amplification, two different primers are used. The first one amplifying only 440bp region of control, whereas, the second primer is able to amplify almost 1100 bp. There is then two band expected in the agarose gel. The ladder in the gel is used to identify the bp as they will show bands at different base pair. Here band size of some individual is same whereas, for some it is different. We can expect a band corresponding to 400 bp region of the ladder and another one corresponding to 1100 bp according to the DNA ladder. Here the PCR product- A obtained from alexix and anna are expected, whereas we can not see any product in case of Briteney and Wan ( A). PCR product - B obtained from Alexis, Anna, Britenex or Wan on the other hand showing the same banding pattern. One band is more than 1000 bp position and second lower band might be coming due to non specific binding of the primer.
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