Recombinant DNA and Genomics

Question 1

recombinant DNA is DNA that is created by

Answer 1

joining together pieces of DNA from various sources

Question 2

recombinant DNA technology is also called

Answer 2

gene splicing

Question 3

clones are

Answer 3

recovered copies of recombinant DNA molecule

Question 4

clones are used to

Answer 4

study structure and orientation of DNA

Question 5

two important tools to construct and amplify DNA molecules are

Answer 5

restriction enzymes and cloning vectors

Question 6

steps to recombinant DNA

Answer 6

1. Cut DNA of interest and vector with restriction enzyme.
2. Incubate together (or cut together).
3. Anneal cut DNA (hydrogen bonds).
4. Ligate nicks in DNA backbone (covalent bonds).
5. Transform vector into host.
6. Screen for host with appropriate vector.
7. Do rad science

Question 7

restriction enzymes cut DNA in what types of ends

Answer 7

stick (cohesive) and blunt ends

Question 8

what is a restrictive enzyme that has been identified in e-coli?

Answer 8

ecoR1

Question 9

DNA ligase

Answer 9

joins fragments sealing the phosphodiester backbone

Question 10

cloning vectors are

Answer 10

DNA molecules that accept DNA fragments

Question 11

distinguishable characteristics of cloning vectors

Answer 11

restriction enzyme sites

selectable gene markers

Question 12

restriction enzyme sites allow

Answer 12

insertion of DNA fragments

Question 13

the selectable gene marker is used to distinguish

Answer 13

host cells that have taken up the DNA fragments from those that have not

Question 14

a successful cloning vector includes

Answer 14

• antibiotic resistance gene intact
• origins of replication intact
• impacted reporter gene

Question 15

plasmids are introduced into bacteria via

Answer 15

transformation

Question 16

transformation uses two main techniques which are

Answer 16

1. calcium ions and a heat shock to pulse DNA into cell

2. electroporation (high intensity pulse)

Question 17

what is the blue-white screening mechanism?

Answer 17

• agar plates containing ampicillin

- agar plates contain x-gal

Question 18

x-gal is blue when

Answer 18

bacterial cells with functional lacZ gene is carrying a nonrecombined plasmid

Question 19

x-gal is white when

Answer 19

bacterial cells contain recombinant plasmid

Question 20

what type of vectors are used to clone large fragments of DNA

Answer 20

BACs an YACs

Question 21

the polymerase chain reaction is a

Answer 21

rapid method of DNA cloning

Question 22

the polymerase chain reaction extends power

Answer 22

of recombinant DNA

Question 23

PCR amplification requires r

Answer 23

• double stranded target DNA
• DNA polymerase
• Mg2+
• four deoxyribonucleoside triphosphates
• primers
• exponential increase of DNA molecule

Question 24

the 3 steps of PCR are

Answer 24

1. denaturation
2. hybridization/annealing
3. extension

Question 25

a limitation of PCR is that it cannot

Answer 25

amplify long segments of DNA

Question 26

what is known as the methodology for studying gene expression?

Answer 26

reverse transcription PCR

Question 27

what is known as the method used to separate DNA fragments by size?

Answer 27

agarose gel electrophoresis

Question 28

DNA sequencing has been through

Answer 28

computer-automated Sanger reaction-based technology

Question 29

DNA sequencing enabled rapid progress of the

Answer 29

human genome project

Question 30

what was discovered as a genome editing tool by scientists studying how bacteria fight viral infections?

Answer 30

CRISPR-Cas

Question 31

CRISPR-Cas system uses

Answer 31

nuclease Cas9

Question 32

CRISPR-Cas stands for

Answer 32

clustered regularly interspaced palendromic repeats

Question 33

CRISPR-Cas takes advantage of

Answer 33

nonhomologous end-joining and nomology-directed repair mechanisms

Question 34

with CRISPR-Cas cas9 or singleguideRNA are introduced into the cell to either

Answer 34

disrupt gene function or create nonfunctional alleles