You are tasked with running a genetic restriction fragment length polymorphism (RFLP) test for the mutant haemachromatos
Posted: Wed May 18, 2022 1:59 pm
You are tasked with running a genetic restriction fragment
length polymorphism (RFLP) test for the mutant haemachromatosis
C282Y allele. Total genomic DNA is purified from the individual to
be tested, the PCR designed in Section B is conducted to amplify
the target region, the 400 bp amplicon is digested with the
restriction enzyme RsaI, and the resulting fragments are analysed
by agarose gel electrophoresis. NB: this particular test depends on
the coincidence that the nucleotide substitution responsible for
the C282Y mutation happens to occur in such a way as to change a
restriction enzyme site, the presence of the RsaI site can be used
a genetic marker for the allele. a) Provide the recognition site
and cleavage position of RsaI in dsDNA. What type of ends are
generated by this enzyme?
length polymorphism (RFLP) test for the mutant haemachromatosis
C282Y allele. Total genomic DNA is purified from the individual to
be tested, the PCR designed in Section B is conducted to amplify
the target region, the 400 bp amplicon is digested with the
restriction enzyme RsaI, and the resulting fragments are analysed
by agarose gel electrophoresis. NB: this particular test depends on
the coincidence that the nucleotide substitution responsible for
the C282Y mutation happens to occur in such a way as to change a
restriction enzyme site, the presence of the RsaI site can be used
a genetic marker for the allele. a) Provide the recognition site
and cleavage position of RsaI in dsDNA. What type of ends are
generated by this enzyme?