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Microbiology > Lecture 9 Biotechnology > Flashcards

Lecture 9 Biotechnology Flashcards

(104 cards)

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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
How is genetically engineered bacteria used in research
Clone DNA segment into bacterium allows easier production of DNA for study Fewer genes in bacterial cell
26
A plant or animal with a cloned gene is ______
transgenic
27
What is DNA library
Collection of clones that contain the entire genome together
28
How do you create a DNA library
Cut DNA with restriction enzyme clone all fragments into population of E.Coli
29
Before cloning eukaryotic genes into bacteria, ______ must be removed
Introns
30
How does a researcher obtain a bacteria without introns
Isolate mRNA reverse transcribe to single DNA strand Synthesize complementary strand to form double stranded cDNA
31
When obtaining bacteria without introns, the complementary strand is synthesized to form double stranded ______
cDNA
32
When generating a recombinant DNA, vector is usually a modified ______
plasmid or bacteriophage
33
the vector has origin of replications and carries ______
Cloned DNA
34
The vector must have ______ sites where DNA is cut so that insert can be joined to it
restriction
35
______ site recognizes insert cut
multiple cloning
36
Vector has ______ marker
selectable - only cells that have vector sequences will grow
37
Second marker is disrupted by ______ when insert is present
Insertional inactivation
38
What does insertional inactivation do
distinguishes recombinant plasmids from intact vector
39
vector ______ selects for cells with vector and differentiates those with recombinant plasmids
pUC18
40
What is the selectable marker
Ampicillin resistance
41
What is the second genetic marker
lacZ' gene
42
What does the product of the lacZ' gene do?
cleaves X-gal, forms blue compound
43
What do the intact vectors become?
blue colonies
44
What do the recombinant molecules become?
White colonies
45
What does DNA sequencing do?
Determines the order of nucleotides in a DNA molecules
46
What type of synteshis is the Dideoxy chain termination method?
In vitro DNA synthesis
47
What does the dideoxy chain termination method require?
Template DNA, DNA polymerase, Primer, neoxynucleotides, dideoxynucleotides
48
in dideoxy chain termination, ddNTP act as ________
Chain terminators
49
What does ddNTP lack
3'OH
50
Synthesis stops when ________ is incorporated
ddNTP
51
in Dideoxy chain termination, different ddNTPs carry ________
Distinct fluorescent marker
52
in Dideoxy chain termination, laser reads ________ labels on separated ddNTPs
fluorescent
53
how is intensity shown by the laser in DCT?
Recorded as a peak
54
In DCT, the order of the color peaks reflect what?
Nucleotide sequence of DNA
55
What is high-throughput sequencing?
high automated methods that generate huge amounts of DNA sequence quickly
56
What does high-throughput sequencing use to form result?
Millions of overlapping small sequences
57
What aligns and merges the data for analysis in high-throughput sequencing?
Computers
58
What does nanopore sequence measure?
brief changes in electrical current to determine nucleotide sequence of long fragments as it passes through microscopic pores in membrane
59
in nanopore sequencing, long fragments of ________ are in electrically conductive solution
ssDNA
60
Nonopore sequencing is when long fragments pass through microscopic pores where different nucleotides ________ electrical currents to varying degrees
block
61
In nanopore sequencing, recorded ________ reflect nucleotide sequence
disruption
62
What sequencing method is used to sequence microbial DNA on international space station?
Nanopore
63
Polymerase Chain Reaction allows for how many DNA?
Creation of over a billion copies of DNA in hours
64
PCR product can be visualized through ________
Gel electrophoresis
65
PCR can detect ________ without culturing
Pathogens
66
What is RT-PCR
Reverse transcription PCR
67
RT-PCR is used to synthesize ________ from mRNA template within a sample
cDNA
68
What is cDNA template for
Amplification
69
What is qPCR
Quantitative PCR
70
qPCR labels the PCR product with ________
Fluorescent
71
Why does qPCR mark the PCR with fluorescent marker?
It's accumulation can be monitored by the marker, which is shown but the amount of fluorescence
72
how does qPCr track the amplification?
in real time
73
What is qPCR primarily used for
Amplification
74
qPCR determines what?
The amount of target DNA in a sample
75
The process of DNA synthesis requires a ________ stranded DNA with a ________ DNA
double, template
76
The process of DNA synthesis requires ________ polymerase
Taq
77
What is Taq polymerase
heat-stable DNA polymerase from Termus aquaticus
78
Process of DNA synthesis requires ________ to determine which portion of the template DNA will be amplified
Primers
79
The process of DNA synthesis requires ________
Deoxynucleotides
80
What are the 4 deoxynucleotides used in DNA synthesis
dATP, dGTP, dCTP, dTTP
81
What is the first step of the amplification cycle in PCR
DNA is denatured by heating, approx. 95 degrees
82
What is the second step of amplification cycle of PCR
Temperature lowered to allow primers to anneal
83
What is the third step of amplification of PCR
Temperature raised to allow DNA synthesis
84
In amplification cycle, target DNA is ________ in each cycle
Doubled, exponential cycle
85
In amplification cycle, what is an important enzyme and why?
Taq polymerase because it is heat-stable and doesn't denature in the step that DNA does
86
________ produce PCR product containing only target sequence
Primers
87
Primers produce PCR product with only ________
Target DNA
88
Primers require how many cycles to produce PCR product?
3
89
What does the sequence of primers determine in PCR?
The length of DNA amplified
90
in PCR, there is ________ amplification of DNA
Exponential
91
in PCR, there is ________ numbers of product of correctly defined length
increasing
92
What are STRs?
Short tandem repeats
93
What does STRs do?
Amplifies certain chromosomal regions
94
STRs repeat within
intron or untranslated region
95
What does the CODIS database contain
Amplification patters of 20 different STR loci (Chromosomal locations) and allows unique identification
96
What do DNA probes do?
Locate specific nucleotide sequence
97
Probe is a ________ DNA piece that is ________ to sequence of interest
single stranded, complementary
98
DNA probes anneal to complement through ________
hybridization
99
the complement and hybridization of DNA probe is labeled with ________
Marker
100
the complement and hybridization of DNA probe is labeled with ________
Marker
101
Colony blotting is used to identify which which ________ contain a sequence of interest
Clones
102
What is fluorescent in situ hybridization
fluorescently labeled probe is used to detect nucleotide sequences in intact cells fixed to microscope slides
103
In FISH, cells are visualized using ________ microscopy
Fluorescence
104
in FISH, probe binds to ________ sequences present in high copy number
RNA

Microbiology (16 decks)

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