SAN DIEGO MESA COLLEGE
Introduction to Microbiology Laboratory (Bio205)
Instructor: Elmar Schmid, Ph.D.
Lab Summary Questions:
________________________
(Student Name)
Q. 1: PCR is an abbreviation and stands for:
P ________________ C _______________ R _________________
Q. 2: Which enzyme is used in PCR to amplify the nucleotide sequence of a template
DNA?
A) DNA polymerase
B) RNA polymerase
C) DNAase
D) Taq polymerase
E) Topoisomerase
Q. 3: You start out with 5 molecules of template DNA and perform 10 PCR cycles with it
(including of course all necessary PCR components). How many DNA molecules will
you have in your PCR reaction tube after completion of the cycles?
A) 15
B) 50
C) 1024
D) 5120
E) 49 million
Q. 4: Which of the following co-factors is required by the polymerase used in PCR
experiments to be catalytically active?
A) Calcium (Ca2+)
B) Manganese (Mn2+)
C) Magnesium (Mg2+)
D) Selenium (Se)
E) Iron (Fe2+)
Q. 5: You have a primer with following nucleotide sequence (see below). What would be
the calculated melting temperature Tm for this primer?
(see formulas posted on the Microbiology part of my website to find the answer!)
5’ – AGTCCGTGACACTCTCATTGGCAC – 3’
Calculated Tm = _________ oC
Q. 6: You would use this primer together with another primer with a Tm of 61.2oC to
perform a PCR experiment. Which annealing temperature would you chose and
program into the thermal cycler?
Annealing temperature TA = ________ oC
Q. 7: What is the advantage of using the HotStart method in PCR DNA amplification
experiments? How does it work?
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SAN DIEGO MESA COLLEGE
Introduction to Microbiology Laboratory (Bio205)
Instructor: Elmar Schmid, Ph.D.
Q. 8: Which of the following components and/or molecules are embedded into the
Ready-To-Go PCR beads you used in this PCR lab and do NOT have to be added to the
PCR reaction mixes? (Make check marks to indicate the presence in the beads)
Ο
Ο
Ο
Ο
Ο
Ο
Ο
Distilled, DNAse-free water
Taq polymerase
forward and reverse primer
Tris buffer
Magnesium chloride (1.5mM)
Non-ionic detergent, e.g. Tween-20
Nucleotides (dNTPs)
Q. 9: For which of the following genes and/or genomic sequences would you expect to
see PCR amplification products even with a small amount of template DNA as 0.5ng
after 30 cycles?
A) Actin gene
B) Hexokinase gene
C) 16S rRNA gene
D) SINE/Alu sequences
E) both c and d
Q. 10: What function has the overlay of PCR reaction mixes with mineral oil (in older
thermal cyclers) or the heated lid in newer generations of thermal cyclers?
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