BIO 351 MIDTERM TEST QUESTIONS AND
ANSWERS
Polymerase Chain Reaction - ANSWER Need sample, Taq polymerase,
nucleotides, primers.
Denature (94-95 C), Anneal (50-56 C), Extend (72 C), then repeat 30x.
DNA Replication - ANSWER Semi-conservative process which produces two
identical strands of DNA.
DNA Pol 1 and 3, SSBPs, Helicase, Ligase, Topoisomerase, Primase, Sliding
Clamp, Leading and Lagging strands, Okazaki fragments
Sanger Sequencing - ANSWER Requires sample, primers, dNTPs, ddNTPs,
and polymerase.
Put four tubes in, replication occurs in each tube-put in the ddNTPs and
replication stops at all different locations of each letter. Use Capillary Gel
Electrophoresis to séquence the DNA by using a laser and detector to detect the
fluorescence of each strand's 3' nucleotide-the ddNTP-from shortest strand to
longest. Result is a graph of peaks of different colors.
Single Molecule Real Time (SMRT) Sequencing ANSWER A DNA sequencing
technology in which nucleotide additions are detected as flashes of fluorescent
colored light, with sensitivity enhanced so that long DNA molecules (up to
15,000 nucleotides) can be sequenced and the results displayed in real time.
Helicase is important and DNA Pol anchors the strand to the bottom of the well
and as nucleotides are added, their fluorescent markers are removed and sensed.
ANSWERS
Polymerase Chain Reaction - ANSWER Need sample, Taq polymerase,
nucleotides, primers.
Denature (94-95 C), Anneal (50-56 C), Extend (72 C), then repeat 30x.
DNA Replication - ANSWER Semi-conservative process which produces two
identical strands of DNA.
DNA Pol 1 and 3, SSBPs, Helicase, Ligase, Topoisomerase, Primase, Sliding
Clamp, Leading and Lagging strands, Okazaki fragments
Sanger Sequencing - ANSWER Requires sample, primers, dNTPs, ddNTPs,
and polymerase.
Put four tubes in, replication occurs in each tube-put in the ddNTPs and
replication stops at all different locations of each letter. Use Capillary Gel
Electrophoresis to séquence the DNA by using a laser and detector to detect the
fluorescence of each strand's 3' nucleotide-the ddNTP-from shortest strand to
longest. Result is a graph of peaks of different colors.
Single Molecule Real Time (SMRT) Sequencing ANSWER A DNA sequencing
technology in which nucleotide additions are detected as flashes of fluorescent
colored light, with sensitivity enhanced so that long DNA molecules (up to
15,000 nucleotides) can be sequenced and the results displayed in real time.
Helicase is important and DNA Pol anchors the strand to the bottom of the well
and as nucleotides are added, their fluorescent markers are removed and sensed.
