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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
Q

The concentration for these DNA and RNA is not in ____ but in _____

A

not in molarity but in mass/volume

26
Q

why is concentration for DNA and RNA expressed in micrograms/mL?

A

small quantities of DNA are used in molecular biology experiments and the size (nucleotides) of the DNA fragments vary

27
Q

Poor lab techniques during DNA isolation can yield DNA samples containing ____,____, and ____

A

Proteins, salts, and ethanol

28
Q

A common practice in molecular biology is to perform a quick assessment of the _____ of nucleic acid samples

A

PURITY

29
Q

How can the purity of DNA be tested?

A

By measuring the absorbance at 260nm and 280nm

Because proteins have a lambda max at 280nm and also some absorbance at 260nm

30
Q

Both DNA and RNA are _______ and have the same ____

A

Both DNA and RNA are COLORLESS and have the same LAMBDA MAX

31
Q

In general, both DNA and RNA solutions have an OD260/OD280 ratio in the range of ___ to ___

A

1.8-2

32
Q

Can the presence of RNA in a DNA sample be accurately detected in a spectrophometric analysis?

A

NO because both DNA and RNA are colorless and have the same lambda max

33
Q

Since differentiating DNA from RNA is not possible in a spectrophometric analysis, what other method can be used to differentiate RNA from DNA?

A

RNA is smaller and migrates faster to the gel - can be detected using agarose gel electrophoresis

34
Q

Purity indication for DNA: using a 260/280 ratio…..

1.8-2 indicates…..
<1.8 indicates…..
>2 indicates……

A

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
Q

Purity indication for DNA: 260/230 ratio

2.0-2.2 indicates……
<2 indicates…….

A

2.0-2.2 indicates PURE DNA
<2 indicates EDTA, CARBOHYDRATES, AND PHENOL

36
Q

What do restriction enzymes do?

A

Cut the DNA at a specific palindromic sequence of base pairs

37
Q

This palindromic sequence of DNA is called the…..

A

Restriction site

38
Q

A palindromic sequence refers to a DNA or RNA segment that has the same sequence when read….

A

3 prime to 5 prime AND 5 prime to 3 prime

39
Q

How are restriction enzymes named?

A

According to the bacterial strain from which the enzyme was first isolated

40
Q

Restriction enzymes are important in ____ and ____

A

DNA analysis and DNA recombinant technology

41
Q

EcroRI was the first enzyme isolated from…

A

Escherichia coli RY13 strain

42
Q

HINDIII was the 3rd enzyme isolated from ____ strain

A

Haemophilius influenzae Rd strain

43
Q

DNA restriction fragments can be cloned into ______ (such as ____) to prepare……

A

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
Q

Restriction digests are also useful to identify ____ in DNA sequences

A

Mutations

45
Q

PGLO plasmid is a circular DNA consisting of ______ base pairs. This plasmid has several restriction enzyme recognition sites including…..

A

5371 base pairs

Eco Ri and HindIII

46
Q

What is a codon?

A

A 3 nucleotide sequence of RNA that codes for a particular amino acid

47
Q

Codons are ______. What does this mean

A

Degenerative, meaning that different codons can translate to the same amino acid

48
Q

What is Mutagenesis?

A

The process by which a change in the nucleotide occurs in a DNA sequence

49
Q

What is a silent mutation?

A

Nucleotide mutation in which without phenotypic, or amino acid, change

50
Q

If a silent mutation occurs within the EcoRi restriction enzyme site, will the mutated sequence be cut by the EcoRi enzyme?

A

NO

51
Q

Since the mutated sequence is not cut by the EcoRi enzyme, how could the differences be seen?

A

During DNA electrophoresis. Different fragments would be seen on the gel

52
Q

What is lambdaDNA?

A

Duplex DNA isolated from bacteriophage

53
Q

Lambda DNA is ____ base pairs in length

A

48,502

54
Q

The sequence of lambdaDNA is commonly used as…

A

As DNA substrate for restriction enzyme

55
Q

Lambda DNA has ___ restriction sites

A

HindIII

56
Q

A HindIII digest of lambda DNA is used as…

A

A DNA marker to determine the size of unknown DNA fragments

57
Q

DNA is a __ ___ but can ___ and ___ itself into more complex shapes

A

DNA is a DOUBLE HELIX but can FOLD and COIL itself into more complex shapes

58
Q

When coiled, DNA size is very ___ and ____

A

Small and compact

59
Q

DNA do most organisms is….

A

Negatively supercoiled

60
Q

When may DNA become nicked?

A

During the extraction process. Creates hooks in the circular DNA

61
Q

Nucleic acid quantification is based on what law?

A

Beer lambert’s law

Biomedical Lab (10 decks)

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