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Microbiology > Lecture 9 Biotechnology > Flashcards

Lecture 9 Biotechnology Flashcards

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1
Q

What is polymerase chain reaction

A

The presence of a DNA segment is detected and the size is determined

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2
Q

What does colony blotting do?

A

Detects specific DNA sequence in colonies grown on agar plates

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3
Q

What cuts DNA strands at recognition sequence

A

Restriction

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4
Q

Restriction enzymes generate ______

A

restriction fragments

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5
Q

Complementary sticky ends can ______

A

Anneal

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6
Q

What does anneal mean

A

Form base pairs with one another

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7
Q

DNA ______ joins molecules covalently

A

ligase

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8
Q

Restriction enzymes allow creation of ______ DNA molecules

A

Recombinant

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9
Q

Restriction enzymes recognize a ______ to ______ base pair nucleotide

A

4 - to 6-

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10
Q

palindromes

A

The same on both strands when read in 5’ to 3’

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11
Q

What does gel electrophoresis do

A

Separates DNA fragments by size

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12
Q

What does the current in the electrophoresis do/

A

causes DNA to migrate through gel to + electrode

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13
Q

In gel electrophoresis, smaller fragments move ______

A

faster

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14
Q

What does CRISPR system do

A

Locates and alters specific sites in DNA molecules

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15
Q

What is the DNA cutting enzyme and what is it used for

A

cas9, gene editing

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16
Q

How does cas9 cut DNA?

A

uses single stranded RNA to recognize DNA

Makes double stranded cut at DNA

DNA can be altered by manipulation DNA repair mechanism

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17
Q

dcas9 (dead) does not cut DNA but still ______

A

uses RNA guide to bind to specific DNA sequence

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18
Q

dCas9 may block ______

A

RNA polymerase

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19
Q

dCas 9 may deliver molecules to certain ______ locations

A

Chromosome

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20
Q

dCas9 can deliver a ______ marker to locate a gene on chromosome

A

Fluorescent

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21
Q

What is genetically engineered bacteria useful for

A

Protein production, DNA production, Research tools

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22
Q

CRISPR cas9 can genetically engineer bacteria, but most engineering uses ______

A

DNA cloning

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23
Q

How does DNA cloning work

isolate DNA
cut with ______ enzyme
join DNA fragment with ______ to generate recombinant molecules
introduce into host where it will ______
high copy number vector makes large amounts of ______

A

restriction
vector (plasmid)
replicate
protein

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24
Q

Why is genetically engineered bacteria better than other sources of bacteria?

A

Safer than taking from animal, won’t cause allergic reactions

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25
Q

How is genetically engineered bacteria used in research

A

Clone DNA segment into bacterium
allows easier production of DNA for study
Fewer genes in bacterial cell

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26
Q

A plant or animal with a cloned gene is ______

A

transgenic

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27
Q

What is DNA library

A

Collection of clones that contain the entire genome together

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28
Q

How do you create a DNA library

A

Cut DNA with restriction enzyme
clone all fragments into population of E.Coli

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29
Q

Before cloning eukaryotic genes into bacteria, ______ must be removed

A

Introns

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30
Q

How does a researcher obtain a bacteria without introns

A

Isolate mRNA
reverse transcribe to single DNA strand
Synthesize complementary strand to form double stranded cDNA

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31
Q

When obtaining bacteria without introns, the complementary strand is synthesized to form double stranded ______

A

cDNA

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32
Q

When generating a recombinant DNA, vector is usually a modified ______

A

plasmid or bacteriophage

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33
Q

the vector has origin of replications and carries ______

A

Cloned DNA

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34
Q

The vector must have ______ sites where DNA is cut so that insert can be joined to it

A

restriction

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35
Q

______ site recognizes insert cut

A

multiple cloning

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36
Q

Vector has ______ marker

A

selectable
- only cells that have vector sequences will grow

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37
Q

Second marker is disrupted by ______ when insert is present

A

Insertional inactivation

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38
Q

What does insertional inactivation do

A

distinguishes recombinant plasmids from intact vector

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39
Q

vector ______ selects for cells with vector and differentiates those with recombinant plasmids

A

pUC18

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40
Q

What is the selectable marker

A

Ampicillin resistance

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41
Q

What is the second genetic marker

A

lacZ’ gene

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42
Q

What does the product of the lacZ’ gene do?

A

cleaves X-gal, forms blue compound

43
Q

What do the intact vectors become?

A

blue colonies

44
Q

What do the recombinant molecules become?

A

White colonies

45
Q

What does DNA sequencing do?

A

Determines the order of nucleotides in a DNA molecules

46
Q

What type of synteshis is the Dideoxy chain termination method?

A

In vitro DNA synthesis

47
Q

What does the dideoxy chain termination method require?

A

Template DNA, DNA polymerase, Primer, neoxynucleotides, dideoxynucleotides

48
Q

in dideoxy chain termination, ddNTP act as ________

A

Chain terminators

49
Q

What does ddNTP lack

A

3’OH

50
Q

Synthesis stops when ________ is incorporated

A

ddNTP

51
Q

in Dideoxy chain termination, different ddNTPs carry ________

A

Distinct fluorescent marker

52
Q

in Dideoxy chain termination, laser reads ________ labels on separated ddNTPs

A

fluorescent

53
Q

how is intensity shown by the laser in DCT?

A

Recorded as a peak

54
Q

In DCT, the order of the color peaks reflect what?

A

Nucleotide sequence of DNA

55
Q

What is high-throughput sequencing?

A

high automated methods that generate huge amounts of DNA sequence quickly

56
Q

What does high-throughput sequencing use to form result?

A

Millions of overlapping small sequences

57
Q

What aligns and merges the data for analysis in high-throughput sequencing?

A

Computers

58
Q

What does nanopore sequence measure?

A

brief changes in electrical current to determine nucleotide sequence of long fragments as it passes through microscopic pores in membrane

59
Q

in nanopore sequencing, long fragments of ________ are in electrically conductive solution

A

ssDNA

60
Q

Nonopore sequencing is when long fragments pass through microscopic pores where different nucleotides ________ electrical currents to varying degrees

A

block

61
Q

In nanopore sequencing, recorded ________ reflect nucleotide sequence

A

disruption

62
Q

What sequencing method is used to sequence microbial DNA on international space station?

A

Nanopore

63
Q

Polymerase Chain Reaction allows for how many DNA?

A

Creation of over a billion copies of DNA in hours

64
Q

PCR product can be visualized through ________

A

Gel electrophoresis

65
Q

PCR can detect ________ without culturing

A

Pathogens

66
Q

What is RT-PCR

A

Reverse transcription PCR

67
Q

RT-PCR is used to synthesize ________ from mRNA template within a sample

A

cDNA

68
Q

What is cDNA template for

A

Amplification

69
Q

What is qPCR

A

Quantitative PCR

70
Q

qPCR labels the PCR product with ________

A

Fluorescent

71
Q

Why does qPCR mark the PCR with fluorescent marker?

A

It’s accumulation can be monitored by the marker, which is shown but the amount of fluorescence

72
Q

how does qPCr track the amplification?

A

in real time

73
Q

What is qPCR primarily used for

A

Amplification

74
Q

qPCR determines what?

A

The amount of target DNA in a sample

75
Q

The process of DNA synthesis requires a ________ stranded DNA with a ________ DNA

A

double, template

76
Q

The process of DNA synthesis requires ________ polymerase

A

Taq

77
Q

What is Taq polymerase

A

heat-stable DNA polymerase from Termus aquaticus

78
Q

Process of DNA synthesis requires ________ to determine which portion of the template DNA will be amplified

A

Primers

79
Q

The process of DNA synthesis requires ________

A

Deoxynucleotides

80
Q

What are the 4 deoxynucleotides used in DNA synthesis

A

dATP, dGTP, dCTP, dTTP

81
Q

What is the first step of the amplification cycle in PCR

A

DNA is denatured by heating, approx. 95 degrees

82
Q

What is the second step of amplification cycle of PCR

A

Temperature lowered to allow primers to anneal

83
Q

What is the third step of amplification of PCR

A

Temperature raised to allow DNA synthesis

84
Q

In amplification cycle, target DNA is ________ in each cycle

A

Doubled, exponential cycle

85
Q

In amplification cycle, what is an important enzyme and why?

A

Taq polymerase because it is heat-stable and doesn’t denature in the step that DNA does

86
Q

________ produce PCR product containing only target sequence

A

Primers

87
Q

Primers produce PCR product with only ________

A

Target DNA

88
Q

Primers require how many cycles to produce PCR product?

A

3

89
Q

What does the sequence of primers determine in PCR?

A

The length of DNA amplified

90
Q

in PCR, there is ________ amplification of DNA

A

Exponential

91
Q

in PCR, there is ________ numbers of product of correctly defined length

A

increasing

92
Q

What are STRs?

A

Short tandem repeats

93
Q

What does STRs do?

A

Amplifies certain chromosomal regions

94
Q

STRs repeat within

A

intron or untranslated region

95
Q

What does the CODIS database contain

A

Amplification patters of 20 different STR loci (Chromosomal locations) and allows unique identification

96
Q

What do DNA probes do?

A

Locate specific nucleotide sequence

97
Q

Probe is a ________ DNA piece that is ________ to sequence of interest

A

single stranded, complementary

98
Q

DNA probes anneal to complement through ________

A

hybridization

99
Q

the complement and hybridization of DNA probe is labeled with ________

A

Marker

100
Q

the complement and hybridization of DNA probe is labeled with ________

A

Marker

101
Q

Colony blotting is used to identify which which ________ contain a sequence of interest

A

Clones

102
Q

What is fluorescent in situ hybridization

A

fluorescently labeled probe is used to detect nucleotide sequences in intact cells fixed to microscope slides

103
Q

In FISH, cells are visualized using ________ microscopy

A

Fluorescence

104
Q

in FISH, probe binds to ________ sequences present in high copy number

A

RNA

Microbiology (16 decks)

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