MOL BIO LAB L10 (Finals- Sequencing Methods)

Question 1

The first DNA sequencing method to be developed and is still widely used today.

Answer 1

Sanger sequencing

Question 2

Two main types of DNA sequencing methods:

Answer 2

1) Sanger sequencing
2) Next-generation sequencing (NGS)

Question 3

The process of determining the order of nucleotides in a DNA molecule.

Answer 3

DNA sequencing

Question 4

First step of Sequencing by ligation (SBL)

Answer 4

DNA molecule is first fragmented into small pieces.

Question 5

Variety of techniques of NGS

Answer 5

• sequencing by synthesis (SBS)
• sequencing by ligation (SBL)

Question 6

This determines the genetic code

Answer 6

order of nucleotides in a DNA molecule

Question 7

The most common type of NGS method

Answer 7

Sequencing by synthesis (SBS)

Question 8

First step of Sequencing by synthesis (SBS)

Answer 8

The DNA molecule is first attached to a solid surface.

Question 9

These methods have revolutionized the field of genomics, making it possible to sequence entire genomes at a relatively low cost

Answer 9

Next-generation sequencing (NGS)

Question 10

NGS methods typically involve massively __________ the sequencing process

Answer 10

parallelizing

Question 11

Arrange in order the following steps involved in Sequencing by synthesis:

____ A laser is used to image the surface and determine the order of the nucleotides in the DNA molecule.

____ A mixture of nucleotides is added to the surface.

____ The nucleotides will bind to the complementary nucleotides in the DNA molecule.

____ The DNA molecule is first attached to a solid surface.

Answer 11

__4__ A laser is used to image the surface and determine the order of the nucleotides in the
DNA molecule.

__2__ A mixture of nucleotides are added to the surface.

__3__ The nucleotides will bind to the complementary nucleotides in the DNA molecule.

__1__ The DNA molecule is attached to a solid surface.

Question 12

This typically involve massively parallelizing the sequencing process, meaning that many DNA molecules are sequenced simultaneously.

Answer 12

Next-generation sequencing (NGS)

Question 13

In SBL, this reads the order of the nucleotides in the chain.

Answer 13

Sequencing machine

Question 14

In SBS, this is used to image the surface and determine the order of the nucleotides in the DNA molecule.

Answer 14

A laser

Question 15

In SBL, the fragments are ________ together to form a long chain.

Answer 15

ligated

Question 16

These are similar to nucleotides, but they lack a 3’-hydroxyl group

Answer 16

Dideoxynucleotides

Question 17

To perform Sanger sequencing, a DNA sample is amplified using ______ and then incubated with a mixture of ______and ______

Answer 17

PCR; ddNTPs and dNTPs

Question 18

In Sanger sequencing, the DNA fragments are separated by ____________ and the sequence of the DNA molecule is determined by ________ the order of the bands on the gel.

Answer 18

gel electrophoresis; reading

Question 19

Arrange in order the following steps involved in Sequencing by ligation:

____ The fragments are attached to adapters.

____ The fragments are then ligated together to form a long chain.

____ DNA molecule is fragmented into small pieces.

____ The sequencing machine then reads the order of the nucleotides in the chain.

____ The adapters contain sequences that are known to bind to the sequencing machine.

Answer 19

__2__ The fragments are
attached to adapters.

__4__ The fragments are then ligated together to form a long chain.

__1__ DNA molecule is fragmented into small pieces.

__5__ The sequencing machine then reads the order of the nucleotides in the chain.

__3__ The adapters contain sequences that are known to bind to the sequencing machine.

Question 20

Next-generation sequencing (NGS) methods revolutionized DNA sequencing by making it possible to?

Answer 20

sequence large DNA molecules much faster and more cheaply

Question 21

In SBL, the fragments are
attached to?

Answer 21

adapters

Question 22

It is a relatively simple and straightforward method, but it can be time-consuming and expensive to sequence large DNA molecules.

Answer 22

Sanger sequencing

Question 23

NGS is now used in a wide range
of applications, including:

Answer 23

research, diagnostics, and personalized medicine

Question 24

Contain sequences that are known to bind to the sequencing machine.

Answer 24

Adapters

Question 25

A collection of methods that can sequence DNA much faster and more cheaply

Answer 25

Next-generation sequencing (NGS)

Question 26

A chain-termination method that uses dideoxynucleotides (ddNTPs) to terminate DNA synthesis.

Answer 26

Sanger sequencing

Question 27

Works by sequencing the DNA molecule one fragment at a time.

Answer 27

Sequencing by ligation (SBL)

Question 28

This randomly terminates DNA synthesis, resulting in a series of DNA fragments of different lengths.

Answer 28

ddNTPs

Question 29

Many DNA molecules are sequenced simultaneously.

Answer 29

Next-generation sequencing (NGS) methods

Question 30

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

Answer 30

Sequencing by synthesis (SBS)

Question 31

This is necessary for DNA synthesis to continue

Answer 31

3'-hydroxyl group

Question 32

The set of instructions for building and maintaining an organism

Answer 32

Genetic code

Question 33

Enumerate the nucleotides

Answer 33

- adenine (A) - thymine (T) - guanine (G) - cytosine (C)

Question 34

Arrange in order the following steps involved in Sanger sequencing: ____ The ddNTPs will randomly terminate DNA synthesis, resulting in a series of DNA fragments of different lengths. ____ The order of the nucleotides in the DNA molecule is determined by reading the order of the bands on the gel. ____ The DNA sample is amplified using PCR. ____ The DNA fragments are separated by gel electrophoresis. ____ The amplified DNA is incubated with a mixture of ddNTPs and dNTPs.

Answer 34

__3__ The ddNTPs will randomly terminate DNA synthesis, resulting in a series of DNA fragments of different lengths. __5__ The order of the nucleotides in the DNA molecule is determined by reading the order of the bands on the gel. __1__ The DNA sample is amplified using PCR. __4__ The DNA fragments are separated by gel electrophoresis. __2__ The amplified DNA is incubated with a mixture of ddNTPs and dNTPs.