27 points total
Biol203 Fall 2013
Homework 8:
Due in your lab starting Nov 13th and will be grading by your Lab TA.
Using Figure 1Q1: You are testing a series of PCR reactions at different temperatures.
Calculate the Tm of oligo 1 assuming 2 degree Celsius per A or T residues and
4 degree Celsius per G or C residue____54___
a: At that temperature calculated in Q1. Using oligos: 1, 2, 3, and 4.
What size PCR products are expected to form after a PCR reaction with:
1 and 4__238bp___
2 and 4____none____
3 and 4____ none _____
b: Using an annealing temperature of 26 degrees Celsius and letting the
annealing reaction go for a few hours (not much energy available at room
temperature to drive annealing of oligos), what size PCR products are
expected to form after a PCR reaction with:
1 and 4__238bp_____
2 and 4____ none ______
3 and 4__238bp ____
Using Figure 2Q2: Using oligo 1 and 4, what are the expected sizes of the PCR products at the
annealing temperature of oligo 1,when trying to amplify SNP1 and SNP2 in
heterozygous or homozygous mutants?
1 and 4 (Homozygous-SNP1)______ none ______
1 and 4 (Heterozygous-SNP1
238bp _____
1 and 4 (Homozygous-SNP2)______ none _____
1 and 4 (Heterozygous-SNP2)_____ 238bp _____
(use info on oligo 2 from Figure 1)
2 and 4 (Homozygous-SNP1)___ 238bp _____
2 and 4 (Heterozygous-SNP1)___238bp ___
2 and 4 (Homozygous-SNP2)_____ none ______
2 and 4 (Heterozygous-SNP2)____ none ______
(use info on oligo 3 from Figure 1)
3 and 4 (Homozygous-SNP1)_____ none ________
3 and 4 (Heterozygous-SNP1)____ none ________
3 and 4 (Homozygous-SNP2)___ 238bp _______
3 and 4 (Heterozygous-SNP2)___238bp _____
27 points total
Biol203 Fall 2013
Using Figure 3Q3: SNP3 and SNP4 each cause the generation of a BamHI sites.
If you PCR genomic DNA heterozygous for SNP3 with:
oligos, 1 and 4, then what will be the approximate sizes of the fragments after
BamHI digestion?___19bp and 219bp and 238bp___
oligos 2 and 4, then what will be the approximate sizes of the fragments after
BamHI digestion?___none_____ and why_______oligo won’t anneal____________
oligos, 3 and 4 then what will be the approximate sizes of the fragments after
BamHI digestion?____ none____and why___________ oligo won’t anneal _______
If you PCR genomic DNA homozygous for SNP3 with oligos:
1 and 4, then what will be the approximate sizes of the fragments after BamHI
digestion?____19bp and 219bp ____________
If you PCR genomic DNA heterozygous for SNP4 with oligos:
1 and 4, then what will be the approximate sizes of the fragments after BamHI
digestion?___ 2x 119bp and 238bp _____________
If you PCR genomic DNA homozygous for SNP4 with oligos:
1 and 4, then what will be the approximate sizes of the fragments after BamHI
digestion?___ 2x 119bp ___________
27 points total
Biol203 Fall 2013