Lecture 6

Sanger sequencing, also known as the “chain

termination method”, is a method for

determining the nucleotide sequence of DNA.

• True

• False

Sanger sequencing is used to generate every possible

length of DNA up to the full length of the target DNA

• True

• False

PCR is used to duplicate the entire DNA sequence

• True

• False

Three Basic Steps of Automated Sanger Sequencing

1) PCR with fluorescent, chain-terminating ddNTPs

2) Size separation by capillary gel electrophoresis

3) Laser excitation & detection by sequencing machine

• True

• False

In manual Sanger sequencing, all ddNTPs are mixed in a single

reaction, and each of the four dNTPs has a unique fluorescent label.

• True

• False

In automated Sanger sequencing, four PCR reactions are set up,

each with only a single type of ddNTP (ddATP, ddTTP, ddGTP, and ddCTP) mixed in.

• True

• False

the chain-terminated oligonucleotides are

separated by size via gel electrophoresis

• True

• False

the oligonucleotides will be arranged from largest to smallest,

reading the gel from top to bottom.

• True

• False

Functional genomic: the study of what certain DNA

fragments codes for & what DNA fragments code

proteins.

• True

• False

Phylogenetic analysis is the study of the evolutionary

development of a species or a group of organisms or

a particular characteristic of an organism

• True

• False