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Biochem-Lab > Exp 4.1: Nucleic Acids (NA) > Flashcards

Exp 4.1: Nucleic Acids (NA) Flashcards

1
Q

Identify:
repository of genetic information

A

nucleic acids

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2
Q

Enumerate:
2 basic structural forms of nucleic acids

A
  1. deoxyribonucleic acid (DNA)
  2. ribonucleic acid (RNA)
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3
Q

Identify which structural form of NA:
genetic material

A

DNA

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4
Q

Identify which structural form of NA:
provides template for RNA transcription

A

DNA

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5
Q

Identify which structural form of NA:
carriers of genetic information for protein translation

A

RNA

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6
Q

Identify:
biopolymers of nucleotide

A

nucleic acids;
(nucleotides form nucleic acids)

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7
Q

Enumerate:
basic components of nucleic acids

A
  1. pentose sugar
  2. heterocyclic nitrogen bases
  3. phosphate
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8
Q

Identify which basic component of NA:
attached by a N-C glycosidic to the 1’-carbon atom of the sugar component

A

heterocyclic nitrogen base

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9
Q

Identify:
nucleotide’s number of phosphate groups

A

1, 2, or 3

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10
Q

Identify:
reason for acidic and anionic character

A

phosphate groups

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11
Q

Identify:
where phosphate is attached

A

attached to the 5’-carbon atom of the sugar component

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12
Q

Identify:
form the symmetrical backbone of the nucleic acid with the 5’ end of one sugar always linked through a phosphate molecule to the 3’ end of the adjacent sugar

A

sugar-phosphate linkages

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13
Q

Enumerate:
interactions responsible for NA rigid molecular configuration

A
  1. phosphodiester bonds
  2. hydrogen bonds
  3. Van der Waal’s forces
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14
Q

Identify the interaction responsible for NA rigid molecular configuration:
links nucleotides in the polynucleotide chain

A

phosphodiester bonds

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15
Q

Identify the interaction responsible for NA rigid molecular configuration:
involved in the complementary base pairing in DNA (& RNA i.e. DNA: RNA hybrids)

A

hydrogen bonds

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16
Q

Identify the interaction responsible for NA rigid molecular configuration:
base stacking - stacking interactions

A

Van der Waal’s forces (pi-pi complexation)

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17
Q

Identify:
a process of DNA purification from a sample using a combination of physical and chemical methods

A

DNA isolation

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18
Q

Enumerate:
methods used for DNA isolation are dependent on?

A
  1. source of sample
  2. age of sample
  3. size of sample
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19
Q

Identify
DNA isolation’s aim

A

to separate the DNA present in the nucleus of the cell from other cellular components

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20
Q

Identify:
major component of all cells;
DNA source

A

nucleic acid
(5-15% of cell’s dry weight)

21
Q

Identify:
choice of DNA sample

A

tissue containing cells with high nuclear-cytoplasmic volume ratio

22
Q

T/F:
DNA concentration is high in the nucleus

A

true

23
Q

Enumerate:
common DNA sources

A
  1. whole blood
  2. hair
  3. sperm
  4. bones
  5. nails
  6. tissues
  7. blood stains
  8. saliva
  9. buccal swabs
  10. urine
24
Q

Enumerate:
organs rich in nucleic acids

A
  1. spleen
  2. liver
  3. thymus
  4. pancreas
25
Q

T/F:
brain is the best source of DNA

A

false;
thymus

26
Q

T/F:
brain and muscle tissues have lower DNA concentration

A

true

27
Q

T/F:
other DNA sources can be archived tissue samples like frozen blood or tissue exhumed from bones, FFPE

A

true

28
Q

Enumerate:
DNA isolation applications

A
  1. paternity testing
  2. forensics
29
Q

T/F:
It is necessary to isolate DNA because presence of proteins, lipids, polysaccharides and other organic/ inorganic components in the DNA preparation can interfere with DNA analysis method.

A

true

30
Q

T/F:
It is necessary to isolate DNA because presence of other components also reduce the quality of DNA leading to its shorter storage life.

A

true

31
Q

Enumerate:
factors to consider in nucleic acid isolation

A
  1. high content of DNA and RNA
  2. low DNAse and RNAse activity
  3. nucleic acids isolated from source are readily obtainable in pure form
  4. large size and fragile nature of DNA
  5. RNA is less stable (compared to DNA)
32
Q

Enumerate reasons:
why the large size and fragile nature of DNA is considered in NA isolation

A
  1. difficulty in isolating intact and undamaged DNA
  2. high molecular weight and intact DNA vs. sheared or fragmented DNA
33
Q

Enumerate reasons:
why the stability of NA is considered in NA isolation

A

RNA: less stable
1. single-stranded molecule
2. presence of 2–hydroxyl group

34
Q

Enumerate:
experimental (physical & chemical) factors

A
  1. pH: extremes in pH
  2. temperature: heating
  3. ionic strength
  4. cellular conditions
  5. mechanical stress
  6. storage of DNA/ RNA
35
Q

Identify the pH range:
hydrogen bonding between complementary strands in stable between?

A

pH 4-10

36
Q

Identify the pH range:
DNA is hydrolyzed at what pH

A

pH less than 3 or greater than 12

37
Q

Identify the temperature range:
most DNA begins to unwind in the range?

A

80°-90°C

38
Q

Identify the temperature:
phosphodiester bonds and β-N, C-glycosidic bonds are stable up to?

A

100°C

39
Q

Identify:
DNA ionic strength and solubility

A

DNA is most stable in ↑[salt];
insoluble ↓[salt solutions]

40
Q

Explain:
reason behind DNA stability and solubility in salt concentrations

A

salt concentrations of less than 0.1M weaken hydrogen bonding between complementary strands

41
Q

Enumerate mechanisms:
cellular conditions as an experimental factor

A
  1. lysis or disruption of the biological membrane release DNA when the tissue is homogenized
  2. presence of nucleases
42
Q

Identify:
grinding, shaking, stirring, squirting the solution through narrow orifices and other disruptive procedures may cause cleavage (shearing or scission) of the DNA chains

A

mechanical stress

43
Q

T/F:
mechanical stress damages DNA secondary structure but increases DNA length

A

false;
does not damage
reduces DNA length

44
Q

T/F:
Purified DNA is best kept in solution.

A

true

45
Q

Explain:
Why is purified DNA best kept in solution?

A

If precipitated DNA is kept dry, it will undergo denaturation.

46
Q

Enumerate:
general steps in DNA isolation

A
  1. homogenization
  2. dissociation and denaturation of the nucleoprotein (deproteinization)
  3. precipitation
  4. purification of nucleic acid
47
Q

Identify the process:
lysis or disruption of biological membranes

A

homogenization

48
Q

Enumerate:
2 ways of hoogenization

A
  1. mechanical: mincing, grinding, sonication
  2. chemical: detergents & chaotropic agents

Biochem-Lab (8 decks)

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