BIOL2164: Principles of Molecular Biology - DNA Sequencing Principles

What is the primary goal of DNA sequencing?

To determine the genetic distance between two organisms
To determine the precise order of nucleotides in a piece of DNA (correct)
To identify the function of a specific gene
To study the evolution of a particular species

Which of the following sequencing technologies is based on the principle of synthesis?

Illumina sequencing (correct)
Nanopore sequencing
Sanger sequencing
Next-generation sequencing

What is the primary difference between Sanger sequencing and next-generation sequencing?

The type of nucleotides used in the reaction
The number of DNA molecules being sequenced in parallel (correct)
The type of sequencing technology used
The size of the DNA fragments being sequenced

What is the purpose of the WACE learning objectives in the context of DNA sequencing?

To provide a framework for understanding the principles of DNA sequencing (C) Signup and view all the answers

What is the advantage of nanopore sequencing over other sequencing technologies?

It is a portable sequencing technology that can be used in the field (D) Signup and view all the answers

What is the rationale for sequencing DNA?

All of the above (D) Signup and view all the answers

What is the significance of identifying organisms through DNA sequencing?

To identify genes/variants associated with specific phenotypes (B) Signup and view all the answers

What is the primary feature of natural DNA replication that is exploited in biotechnology to develop DNA sequencing techniques?

Complementary base pairing (C) Signup and view all the answers

What is the role of ddNTPs in Sanger sequencing?

To terminate the chain reaction (C) Signup and view all the answers

Why is only one of the two ssDNA strands used for sequencing in Sanger sequencing?

Because only one primer is used (D) Signup and view all the answers

What is the purpose of the 4 reaction tubes in Sanger sequencing?

To terminate the chain reaction with different ddNTPs (A) Signup and view all the answers

What is the result of the Sanger sequencing reaction when the ddATP is added?

5 terminated A fragments are acquired (A) Signup and view all the answers

What is the purpose of polyacrylamide gel electrophoresis in Sanger sequencing?

To separate the DNA fragments based on size (B) Signup and view all the answers

What is the result of the collection of fragments in Sanger sequencing?

A sequencing ladder (A) Signup and view all the answers

Why is autoradiography used in Sanger sequencing?

To visualize the DNA sequence (B) Signup and view all the answers

What is the significance of DNA polymerase in Sanger sequencing?

Not any DNA polymerase can be used for sequencing (D) Signup and view all the answers

What is the primary function of the 5'T>3' exonuclease activity in DNA polymerase I?

To remove mistakes in the 5'T>3' direction (C) Signup and view all the answers

Which of the following DNA polymerases is NOT suitable for sequencing?

Any DNA polymerase (B) Signup and view all the answers

What is the limitation of Klenow polymerase in sequencing?

It has low processivity (A) Signup and view all the answers

What is the main advantage of Taq polymerase in sequencing?

It has high processivity (A) Signup and view all the answers

What is the purpose of the fluorescence-labeled ddNTPs in automated Sanger sequencing?

To emit light of a specific color (A) Signup and view all the answers

What is the limitation of the Sanger sequencing method?

It is limited to sequencing DNA fragments of up to 750 bp (D) Signup and view all the answers

What is the purpose of assembling the sequence in both directions?

To confirm the sequence of the gene (C) Signup and view all the answers

Why is the human genome difficult to sequence using the Sanger method?

It is too large (B) Signup and view all the answers

What is the purpose of the polyacrylamide gel or capillary sequencer in automated Sanger sequencing?

To separate DNA fragments (A) Signup and view all the answers

What is the advantage of Next-Generation Sequencing (NGS) technologies?

They are higher-throughput than Sanger sequencing (B) Signup and view all the answers