Question

Can someone help me with this homework question on gel electrophoresis?

Translocation and deletion 3. You PCR amplify a 500 bp (base pairs) piece of DNA that has diagnostic value in determining whether a patient has a mutation within a specific DNA region. You know that this DNA segment of the "normal" gene does not include an EcoRI restriction site; but the mutated DNA segment of the same gene contains an EcoRI restriction site due to a point mutation at the 100th bp from the 5' DNA end. After PCR amplification, you subject your DNA to EcoRI digestion, and separate both, the undigested and digested DNA samples by gel electrophoresis. a. Draw a schematic of DNA segments for both normal and mutated DNA. (2 points) b. Draw the results of the gel electrophoretic separation after EcoRI digestion for normal (lane 1) and mutated DNA segment (lane 2), assuming the patient carries the mutation only on one parental allele (4 points) Marker 1 2 1000 bp 500 bp 200 bp 100 bp c. Draw the results of the gel electrophoretic separation after EcoRI digestion for normal (lane 1) and mutated DNA segment (lane 2), assuming the patient carries the mutation on both parental alleles (4 points) Marker 1 2 1000 bp 500 bp 200 bp 100 bp

Answer 1

The answer to the given question has been solved as under;

Bour vons : 3 a). ginen original piece of DNA a 500 lehr Normal gene doucnot have restriction. A rxpected sime after degistion 500 h Mutated gine has a restricted site at 100 ah from 5' site . Expected size after digestion - 100 hp + (500-100) ep 100 up + 400 bp. Normal DNA 4 Mutated DNA a 5 500k 31- 3 4 + 35 $ Marku 0 after 100off 500 parental Separation product EcoRI alig istion, mutation on one allele only. - - - - - 11400. JEAboup Roolip- 1001 - - 100 bp. Marker 10004 Separation product after reco RI digestion ; mutation on both harental allites. 50044 - - - - 600up $400 kop 400 bin 2004 - 100 hp -