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Biomedical Lab > Lab 7 > Flashcards

Lab 7 Flashcards

1
Q

The genotype of an organism is determined by….

A

the DNA content that is passed onto the organism by its parents

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2
Q

What is GENOMICS?

A

The study of genes

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3
Q

Typically, individuals of the same species have similarities in their _____, but not all of their _____ are identical

A

individuals of the same species have similarities in their GENOME but not all of their genomic DNA sequences are identical

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4
Q

Even individuals who are very closely related have many different…..

A

genetic differences

exceptions =twins or individuals cloned from the same parent

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5
Q

During bacterial reproduction, the clones contain a direct copy of DNA from the….

A

original parent bacterial cell

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6
Q

a bacterial colony observed on an agar plate contains….

A

millions of bacteria cloned from a single parent bacteria

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7
Q

Is DNA replication always perfect?

A

NO - single nucleotide mutations are common

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8
Q

PHENOTYPE of an organism is determined by…

A

observed physical and behavioral characteristics of an organism

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9
Q

Give some examples of phenotype

A

appearance (shape, size, color)
metabolic activity
movement

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10
Q

What things allow the genome to be expressed as a phenotype?

A

translation of mrna to protein, protein-protein interactions, and availability of cofactors

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11
Q

RNA molecules transcribed from different genes contain…

A

information that specifies the structure of proteins from which cells are built

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12
Q

What is a mutation?

A

a change in the nucleotide sequence of a short region of the genome and results in changes to a genome

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13
Q

In general, do mutations cause phenotypic changes?

A

NO

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14
Q

Under what circumstances WOULD a mutation cause a phenotypic change?

A

If they drastically affect the protein expression (transcription/translation) and/or structure

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15
Q

What are POLYMORPHISMS?

A

mutations that do NOT cause disease and are present in greater than 1% of the population

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16
Q

What are silent mutations?

A

mutations that do not show a change in the translated amino acid and hence no change in phenotype

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17
Q

Polymorphisms are _____ genetic differences among organisms in the same _____

A

polymorphisms are NATURALLY OCCURING genetic differences among organisms in the same SPECIES

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18
Q

The most common type of polymorphism involves…..

A

variation at a single base pair

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19
Q

When is NUCLEIC ACID QUANTIFICATION performed?

A

before performing downstream experiments using the extracted DNA and RNA

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20
Q

State the nucleic acid quantification formula

A

c=A/EL

c=nucleic acid concentration (Molar)

A=UV absorbance in absorbance units (AU)

E=wavelength-dependent molar absorptivity coefficient in M^-1cm^-1

L= Light path in cm

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21
Q

BOTH DNA and RNA are _____ and have a lambda max at ______

A

colorless, 260nm

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22
Q

Why is a standard curve NOT required for DNA and RNA?

A

There is a published average extinction coefficient value for DNA and RNA

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23
Q

What is the average extinction coefficient for double stranded DNA?

A

0.020(microgram/mL)^-1cm^-1

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24
Q

What is the average extinction coefficient for single stranded RNA?

A

0.025(microgram/mL)^-1cm^-1

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25
The concentration for these DNA and RNA is not in ____ but in _____
not in molarity but in mass/volume
26
why is concentration for DNA and RNA expressed in micrograms/mL?
small quantities of DNA are used in molecular biology experiments and the size (nucleotides) of the DNA fragments vary
27
Poor lab techniques during DNA isolation can yield DNA samples containing ____,____, and ____
Proteins, salts, and ethanol
28
A common practice in molecular biology is to perform a quick assessment of the _____ of nucleic acid samples
PURITY
29
How can the purity of DNA be tested?
By measuring the absorbance at 260nm and 280nm Because proteins have a lambda max at 280nm and also some absorbance at 260nm
30
Both DNA and RNA are _______ and have the same ____
Both DNA and RNA are COLORLESS and have the same LAMBDA MAX
31
In general, both DNA and RNA solutions have an OD260/OD280 ratio in the range of ___ to ___
1.8-2
32
Can the presence of RNA in a DNA sample be accurately detected in a spectrophometric analysis?
NO because both DNA and RNA are colorless and have the same lambda max
33
Since differentiating DNA from RNA is not possible in a spectrophometric analysis, what other method can be used to differentiate RNA from DNA?
RNA is smaller and migrates faster to the gel - can be detected using agarose gel electrophoresis
34
Purity indication for DNA: using a 260/280 ratio….. 1.8-2 indicates….. <1.8 indicates….. >2 indicates……
1.8-2 indicates PURE DNA <1.8 indicates PRESENCE OF PROTEIN, ALCOHOL, ACIDIC PH, OR OTHER CONTAMINANTS >2 indicates RNA CONTAMINATION, BASIC PH
35
Purity indication for DNA: 260/230 ratio 2.0-2.2 indicates…… <2 indicates…….
2.0-2.2 indicates PURE DNA <2 indicates EDTA, CARBOHYDRATES, AND PHENOL
36
What do restriction enzymes do?
Cut the DNA at a specific palindromic sequence of base pairs
37
This palindromic sequence of DNA is called the…..
Restriction site
38
A palindromic sequence refers to a DNA or RNA segment that has the same sequence when read….
3 prime to 5 prime AND 5 prime to 3 prime
39
How are restriction enzymes named?
According to the bacterial strain from which the enzyme was first isolated
40
Restriction enzymes are important in ____ and ____
DNA analysis and DNA recombinant technology
41
EcroRI was the first enzyme isolated from…
Escherichia coli RY13 strain
42
HINDIII was the 3rd enzyme isolated from ____ strain
Haemophilius influenzae Rd strain
43
DNA restriction fragments can be cloned into ______ (such as ____) to prepare……
DNA restriction fragments can be cloned into DNA VECTORS (such as PLASMIDS) to prepare RECOMBINANT DNA MOLECULES USED FOR PROTEIN SYNTHESIS AND IN GENE ANALYSIS
44
Restriction digests are also useful to identify ____ in DNA sequences
Mutations
45
PGLO plasmid is a circular DNA consisting of ______ base pairs. This plasmid has several restriction enzyme recognition sites including…..
5371 base pairs Eco Ri and HindIII
46
What is a codon?
A 3 nucleotide sequence of RNA that codes for a particular amino acid
47
Codons are ______. What does this mean
Degenerative, meaning that different codons can translate to the same amino acid
48
What is Mutagenesis?
The process by which a change in the nucleotide occurs in a DNA sequence
49
What is a silent mutation?
Nucleotide mutation in which without phenotypic, or amino acid, change
50
If a silent mutation occurs within the EcoRi restriction enzyme site, will the mutated sequence be cut by the EcoRi enzyme?
NO
51
Since the mutated sequence is not cut by the EcoRi enzyme, how could the differences be seen?
During DNA electrophoresis. Different fragments would be seen on the gel
52
What is lambdaDNA?
Duplex DNA isolated from bacteriophage
53
Lambda DNA is ____ base pairs in length
48,502
54
The sequence of lambdaDNA is commonly used as…
As DNA substrate for restriction enzyme
55
Lambda DNA has ___ restriction sites
HindIII
56
A HindIII digest of lambda DNA is used as…
A DNA marker to determine the size of unknown DNA fragments
57
DNA is a __ ___ but can ___ and ___ itself into more complex shapes
DNA is a DOUBLE HELIX but can FOLD and COIL itself into more complex shapes
58
When coiled, DNA size is very ___ and ____
Small and compact
59
DNA do most organisms is….
Negatively supercoiled
60
When may DNA become nicked?
During the extraction process. Creates hooks in the circular DNA
61
Nucleic acid quantification is based on what law?
Beer lambert’s law

Biomedical Lab (10 decks)

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