24 points total
Biol203 Fall 2013
Homework 7:
Due in your lab starting Oct 30th and will be grading by your Lab TA.
Q1: You will use RFLP analysis for part 1 and 2 to identify the HindIII SNP in
the population.
WT would NOT contain HindIII sites in both alleles.
Homozygous mutant would contain HindIII sites at both alleles.
Make sure that you describe how you would observe in a WT (absence of
HindIII) site, Heterozygote or Homozygous (containing both HindIII sites).
Part 1: 3pts. Draw a picture, labeled appropriately, that shows how you could
detect this mutation by Southern blot hybridizations?
24 points total
Biol203 Fall 2013
Part 2: 3pts. Draw a picture and label the provided genomic region as to how
primers could be used for PCR to detect SNP without the use of sequencing and
what steps you would follow after PCR.
24 points total
Biol203 Fall 2013
Part 3: 4pts. Please describe/draw how you could use oligonucleotide
hybridization on whole genomic DNA or PCR products to detect HindIII
polymporhpism? Be specific.
2pts for getting close to the right protocol
1pt for the WT oligo
1pt for the mutant oligo
or
24 points total
Biol203 Fall 2013
Q2. Using the following restriction digest data from a 6kb plasmid DNA, please
draw a restriction maps for this plasmid.
Remember: (5pts.)
1 cut in a circular plasmid gives__1_ fragment.
2 cuts in a circular plasmid gives __2___fragments.
3 cuts in a circular plasmid gives ___3__fragments.
4 fragments equals how many cuts in a circular plasmid?_ 4__
5 fragments equals how many cuts in a circular plasmid?__ 5__
Sizes of fragments when run on an Agarose gel after 6kb plasmid is cut:
EcoR1
3 kb
2 kb
1 kb
BamHI
3 kb
3 kb
6pts for Bam/EcoR1 map
BamHI/EcoR1
1.75 kb
1.75 kb
1.25 kb
1 kb
0.25 kb
Be AWARE that students may draw the mirror image for
these maps, which is correct as well. 1pt for each site
correctly placed (5pts) and 1pt for the whole thing.
24 points total
Biol203 Fall 2013
Sizes of fragments when run on an Agarose gel after 6kb plasmid is cut:
HindIII
2.5 kb
2 kb
1.5 kb
BamHI/HindIII
1.75 kb
1.5 kb
1.25 kb
1.25 kb
.25 kb
EcoR1/HindIII
2.5 kb
1.5 kb
1 kb
0.5 kb
0.5 kb
0.01 kb
3pts for Bam/EcoR1/HindIII map Be AWARE that students may draw the
mirror image for these maps, which is correct as well. 1pt for each site
correctly placed (3pts).