Based on the DNA extraction of Human Buccal Cells lab experiement with the overa

Question

Based on the DNA extraction of Human Buccal Cells lab experiement with the overall intention of cloning our own DNA , RFLP analysis Electrophoresis( 1.5% agarose) was conducted. Using the PCR A & B RFLP Analysis Gel electrophoresis results given ; Please provide short, concise, and detailed explanation in respect to the movement of DNA through the agarose gel (1.5 %)( i.e. speed/ size/ etc. of fragments, ~size of Base pairs), etc. Also, based on these results obtained from this gel electrophoresis run, please address any possible sources of error we may have encountered for this particular gel run,especially since Student # 5 may have used the same restriction enzyme for PCR tubes A & B.

RFLP Anyalysis-->(Restriction fragment length polymorphism)

IMPORTANT: For Gel Run (RFLP Electrophoresis) PCR Products A & B were cut with RESTRICTION ENZYMES MseI & RsaI.

Also, for this particular part of the experiment Student # 5 may have accidentally used the same restriction enzyme for both PCR tubes A & B as seen in gel ladders 16&17, how did this mistake affect the results of this gel electrophoresis run? Is this why both A & B for ladders 16& 17 are at the same level ? How does this compare to "me!" results and that of other students as well. Please be detailed ty!

RFLP Gel Electrophoresis:

Explanation / Answer

The pcr productA and B of rflp gel electrophoresis is amplified by student 1, 2 and 3 are getting some nonspecific and multiple bands it means the pcr is not effeciently done or primer anealing temperature is less therfore the primer binding could have been nonspecific. Now coming to the size of the pcr A and B product, as the A pcr product run faster than B it means the A has lesser size than B. The ladder size is not given but according to 1kb ladder the pcr A has 300bp size and pcrB has 1500bp size. Nowcoming to the digestion part as given in the question that 5th student used same restrintion enzyme rather than using different two restriction enzymes gives the digested product but cut from the only one end in simple terms only nicked dna is present that is why both the pcr digested product size is same and the movement is also same.

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