DNA sequencing

Question 1

____________ is the process of determining the order of nucleotides in a DNA molecule.

Answer 1

DNA sequencing

Question 2

What are the 4 major Nucleotides?

Answer 2

• Adenine (A)
• Thymine (T)
• Cytosine (C)
• Guanine (G)

Question 3

The order of ___________ in a DNA molecule determines the ___________

Answer 3

nucleotides; genetic code

Question 4

_____________ the set of instructions for building and maintaining an organism.

Answer 4

genetic code

Question 5

What are the two main types of DNA sequencing methods?

Answer 5

• Sanger method
• next-generation sequencing (NGS)

Question 6

chain-termination method

Answer 6

Sanger sequencing

Question 7

Sanger sequencing uses _____________ to terminate DNA synthesis.

Answer 7

dideoxynucleotides (ddNTPs)

Question 8

Dideoxynucleotides are similar to nucleotides, but they lack a ____________ which is necessary for DNA synthesis to continue

Answer 8

3’-hydroxyl group,

Question 9

To perform Sanger sequencing, a DNA sample is amplified using ________ and then incubated with a mixture of ________________ The ddNTPs will randomly terminate DNA synthesis, resulting in a series of DNA fragments of different lengths. The DNA fragments are then separated by ______________ and the sequence of the DNA molecule is determined by reading the order of the bands on the gel.

Answer 9

• PCR
• ddNTPs and dNTPs
• gel electrophoresis

Question 10

_______________ is a collection of methods that can sequence DNA much faster and more cheaply than Sanger sequencing.

Answer 10

Next-generation sequencing

Question 11

What are the advantages of NGS?

Answer 11

sequence much faster & more cheap then Sanger sequencing

Question 12

NGS methods typically involve __________________, meaning that many DNA molecules are sequenced simultaneously

Answer 12

massively parallelizing the sequencing process

Question 13

Two techniques employed in NGS?

Answer 13

• Sequencing-by-synthesis (SBS)
• Sequencing-by-ligation (SBL)

Question 14

NGS methods have revolutionized the _____________ making it possible to sequence entire genomes at a relatively low cos

Answer 14

field of genomics

Question 15

What are examples of wide applications of NGS?

Answer 15

• research,
• diagnostics,
• and personalized medicine

Question 16

the first DNA sequencing method to be developed

Answer 16

Sanger sequencing

Question 17

Give the advantages and disadvantages of Sanger sequencing

Answer 17

Pros:
Relatively simple and Straightforward

Cons:
Time consuming and expensive to sequence large DNA molecules

Question 18

What are the steps involved in Sanger sequencing?

Answer 18

1. The DNA sample is amplified using PCR.
2. The amplified DNA is incubated with a mixture of ddNTPs and dNTPs.
3. The ddNTPs will randomly terminate DNA synthesis, resulting in a series of DNA fragments of different lengths.
4. The DNA fragments are separated by gel electrophoresis.
5. The order of the nucleotides in the DNA molecule is determined by reading the order of the bands on the gel.

Question 19

the most common type of NGS method

Answer 19

Sequencing-by-synthesis

Question 20

how does SBS work?

Answer 20

It works by sequencing the DNA molecule one nucleotide at a time.

Question 21

used to image the surface and determine the order of the nucleotides in the DNA molecule.

Answer 21

laser

Question 22

how does SBL work?

Answer 22

sequencing the DNA molecule one fragment at a time

Question 23

contain sequences that are known to bind to the sequencing machine

Answer 23

adapters

Question 24

reads the order of the nucleotides in the chain.

Answer 24

Sequencing machine