a) Provide the recognition site and cleavage position of Rsal in dsDNA. What type of ends are generated by this enzyme?
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a) Provide the recognition site and cleavage position of Rsal in dsDNA. What type of ends are generated by this enzyme?
a) Provide the recognition site and cleavage position of Rsal in dsDNA. What type of ends are generated by this enzyme? [2] b) The sequence below represents the 400 bp PCR amplicon of the HFE WT allele. Indicate the position and nature of the point mutation responsible for the C282Y mutation. Use a yellow highlight to indicate the potential recognition site(s) for the enzyme Rsal. [2] ggcaagggtaaacagatcccctctcctcatccttcctctttcctgtcaag gectectetggtgaaggtgacacatcatgtgacctcttca gtgaccactctacggtgtcgggccttgaactactacdcccagaacatcaccatgaagtggctgaaggataagcagccaatggatgea aaggagttegaacetaaagacgtattgcecaatggggatgggacctaccagggctggataaccttggctgtacoccetggggaaga gcagagatatacgtgccaggtggagcacccaggcctggatcagcccctcattgtgatotggggtatgtgactgatgagagccagga gctgagaaaatctattgggggttgagaggagtgcctgaggaggtaattat c) Using the line diagram below, indicate where you would fine the Rsal sites on your PCR amplicons for a patient who is heterozygous for the C282Y mutation. Include the fragment sizes. [3] d) You analyse DNA samples with your PCR RFLP assay developed above, followed by agarose gel electrophoresis. Draw the gel banding pattern with sizes you expect to observe for a wild type homozygote, mutant homozygote and heterozygote (carrier) individual respectively. [3]
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