WSSP CH3 DNA Sequencing PS1
Name: ______________________________ School: ________________________
Chapter 3 Problem Set1:
1. (8 points) DNA is made up of two, three base pair, complementary strands of
repeating units of ribose, purine or pyrimidine base, and phosphate. Using the
following symbols…
…link the units together to form a three base pair, double stranded DNA.
Show six (6) ribose units. Show covalent bonds be solid lines and hydrogen
bonds by dotted lines. Prepare your illustration in the space provided below:
WSSP CH3 DNA Sequencing PS1
2. (1 point) How are the DNA chains separated for DNA sequence analysis?
3. (6 points) Can nucleotide A, B and C
be used for chain elongation in
sequence analysis or not? STATE WHY,
or you will not get credit. Give an answer
and explanation for A, for B, and for C.
4. (6 points) Can nucleotide A, B and C
be used to terminate the DNA chain in
sequence analysis or not? STATE WHY,
or you will not get credit. Give an
answer and explanation for A, for B,
and for C.
WSSP CH3 DNA Sequencing PS1
5) Name the two critical features of di-deoxy NTPs that enable DNA
sequence to be determined. (4 points)
6) (8 points) Consider these potential outcomes of mistakes in
sequencing:
1) No sequence at all;
2) Very small fragments;
3) Missing information for one nucleotide;
4) Mostly illegible sequence data
5) Average fragment length becomes shorter
6) Average fragment length becomes longer
7) No apparent sequence but long fragments are synthesized
8) Sequence reaction is fine
Indicate the outcome (1, 2 , 3 etc) for each experiment and describe WHY.
a) You add the PCR forward and reverse primers to your DNA sequencing
reaction.
b) Your miniprep DNA was not eluted from the column.
c) Your plasmid did not include a cDNA insert.
d) You forgot to add the ddNTPs until your lab partner reminded you—so you
added them after the first half hour of the reaction.