DNA Sequencing

Question 1

In manual Sanger DNA sequencing using radioactively labeled primers, how many reaction tubes are necessary?

Answer 1

4 tubes, 1 for each dideoxynucleotide

Question 2

In automated DNA sequencing using fluorescently labeled nucleotides, how many reaction tubes are necessary?

Answer 2

1, all fluorescently labeled nucleotides can be combined in one tube

Question 3

There is a limit to how many basepairs of DNA can be sequenced in any single sequencing reaction. If automated Sanger dideoxy sequencing is used, what is the limit?

Answer 3

1000 bp

Question 4

Which 2 sequencing methods rely on separation of labeled DNA fragments by electrophoresis?

Answer 4

Maxam-Gilbert and Sanger; this limits sequencing long stretches of DNA

Question 5

Which 3 methods rely on DNA synthesis?

Answer 5

Sanger, Pyrosequencing, and Illumina

Question 6

Which 2 methods require making a genomic library in a cloning vector?

Answer 6

Maxam-Gilbert and Sanger

Question 7

What is the main difference between Ion semiconductor sequencing and pyrosequencing?

Answer 7

Ion semiconductor sequencing measures the release of H+ instead of pyrophosphate