Sequencing Technologies BTN602C Genomics and Proteomics

Which part of Sanger (Cycle) Sequencing is NOT found in regular Polymerase Chain Reaction (PCR)?

• DNA polymerase

• Primers

• Deoxyribonucleotides (dNTPs)

• Dideoxyribonucleotides (ddNTPs)

• DNA template

How are deoxyribonucleotides different from dideoxyribonucleotides? Choose all that apply.

• Deoxyribonucleotides are used in both PCR and Cycle Sequencing.

• Dideoxyribonucleotides are missing an OH group compared to deoxyribonucleotides.

• You cannot add new nucleotides to a deoxyribonucleotide.

• Deoxyribonucleotides are NOT found in natural living cells.

• Dideoxyribonucleotides are labeled with fluorescent tags.

How is DNA synthesis in PCR and Cycle Sequencing the SAME?

• Both require DNA polymerase.

• Both require primers.

• Both require deoxynucleotides (dNTPs).

• Both require dideoxynucleotides (dNTPs).

• Both generate new DNA products of equal length.

Place the steps of DNA Sequencing below into their correct order.

A. Detection of fluorescence w/ lasers & sensors

B. PCR w/ fluorescent dideoxynucleotides

C. Separate DNA by size w/ gel electrophoresis

D. Match DNA sequence using BLAST database

• A, B, C, D

• B, C, A, D

• B, A, C, D

• D, B, C, A

Which of the following is chemical nucleotide sequencing method

• Sanger method

• Maxam Gilbert method

• Edmans method

• Automated sequencing method

The principle of Sanger's method relies on

• use of chemicals for base specific cleavage

• use of dNTPs for chain termination

• use of ddNTPs for chain termination

• Use of p32 for chain termination

Automated DNA sequencing is an improvement of Sanger's method where

• ddNTPs are used for chain termination

• PCR is used for making sequencing

• Fluorescently labelled dNTPs are used for chain termination

• Fluorescently labelled ddNTPs are used for chain termination

NGS consists of the following steps in sequence

• library construction, amplification, sequencing

• sequencing, library construction, amplification

• Amplification, Sequencing and library construction

• none of the above

Emulsion PCR refers to

• amplification of DNA in water

• amplification of DNA in organic solvent

• amplification of DNA in water in oil droplets

• synthesis of DNA in water

problems with 454

• repeat regions cant be read

• cant be used for large genomes

• allows in depth sequencing

• short stretches are read