Brainscape's Knowledge GenomeTM


Top Flashcards (Certified)
All Flashcards

MolBio > DNA and RNA extraction > Flashcards

DNA and RNA extraction Flashcards

(49 cards)

Start Studying
1
Q

starting point in any molecular diagnostic kit

A

nucleic acid extraction

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
2
Q

these methods rely on biochemical properties of the cellular components to elicit the desired molecular separation and might exhibit preference or exclusivity in extracting DNA or RNA depending on its intrinsic characteristics

A

NAE chemical method

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
3
Q

enumerate the chemical techniques

A
  • osmotic shock
  • enzymatic digestion
  • detergents
  • alkali treatment
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
4
Q

osmotic rupture of membrane

A

osmotic shock

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
5
Q

principle of enzymatic digestion

A

digestion of cell wall

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
6
Q

principle of detergent

A

solubilization of membranes

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
7
Q

principle of alkali treatment

A

solubilization of membrane

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
8
Q

mode of lysis (chemical)

A
  • osmotic shock - gentle
  • enzymatic digestion - gentle
  • detergents - gentle
  • alkali treatment - harsh
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
9
Q

enzymatic digestion cost

A

cheap at small scale
expensive at large scale

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
10
Q

most usual application of osmotic shock

A

spheroplasts and protoplasts

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
11
Q

most usual application of enzymatic digestion

A

gram-positive and gram-negative bacteria

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
12
Q

most usual application of detergents

A

general use

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
13
Q

most usual application of alkali treatment

A

plasmid DNA

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
14
Q

Cesium Chloride Gradient Centrifugation with Ethidium Bromide is baded on the phenomenon ____

used to ____

A

phenomenon of buoyant and specific density

used to extract DNA from bacteria

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
15
Q

separation of RNA from DNA and proteins after extraction with an acidic solution

A

Guanidinium Thiocyanate-Phenol-Chloroform Extraction

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
16
Q

Guanidinium Thiocyanate-Phenol-Chloroform Extraction consist mainly of?

A
  • GuSCN
  • sodium acetate
  • phenol
  • chloroform

followed by centrifugation

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
17
Q

in Guanidinium Thiocyanate-Phenol-Chloroform Extraction, total RNA remains in the ____ while most of DNA and proteins part remain either in the ____

A

total RNA = upper aqueous phase

DNA and proteins = interphase or in the lower organic phase under acidic condition

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
18
Q

Guanidinium Thiocyanate-Phenol-Chloroform Extraction: total RNA Is then recovered through?

A

precipitation by isopropanol

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
19
Q

Chelex extraction is used in the field of?

A

forensics for DNA extraction from various sources such as:
- hair
- blood stain cards
- buccal swabs

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
20
Q

inhibiting DNA degradation by chelating metal ions which cause DNA breakdown at high temperature and lower ionic conditions

Study These Flashcards
A

Chelex extraction

21
Q

alkaline extraction is dedicated to?

Study These Flashcards
A

plasmid DNA isolation

22
Q

principle of alkaline extraction

Study These Flashcards
A

selective alkaline denaturation of high molecular weight chromosomal DNA, while covalently bond circular plasmid DNA remains intact. After neutralization, chromosomal DNA renatures and makes an insoluble precipitate, while plasmid DNA remains in the supernatant

23
Q

involves harvesting the bacteria of interest from culture media and exposing them to alkaline solution (consisting basically of SDS and NaOH)

Study These Flashcards
A

alkaline extraction

24
Q

poly(A) RNA hybridizes with an oligo(dT)-cellulose matrix, under high-salt conditions

Study These Flashcards
A

purification of Poly(A) + RNA by Olido(dT)-Cellulose chromatography

25
based on the liquid and stationary phases, which **selectively separate the target analyte from the solution** based on specific hydrophobic, polar, and/or ionic properties of both solute and sorbent
Solid-Phase nucleic acid Extraction - normal/regular SPE - reverse SPE - ion exchange SPE
26
solid phases use (6)
- silica matrices - glass particles - diatomaceous earth - magnetic beads-based nucleic acid purification - anion exchange material - cellulose matrix
27
enumerate the mechanical techniques
- homogenization (blade or pestle) - ultrasonication or cavitation - pressure cell "french press" - ball mill
28
method of choice for large scale
homogenization
29
most usual application of homogenization
animal tissues
30
most usual application of ultrasonication or cavitation
good for spheroplasts but not primary cells
31
most usual application of pressure cell
gram negative and some gram positive bacteria
32
most usual application of ball mill
bacteria, yeast, microalgae, unicellular animal cells
33
principle of homogenization
shredding of cells
34
principle of ultrasonication or cavitation
disruption of cells by pressure
35
principle of pressure cell
disruption of cells by shear force
36
principle of ball mill
cells crushed between glass/steel balls/beads
37
most common technique to determine DNA yield and purity?
measurement of absorbance
38
needed for absorbance method
spectrophotometer with UV lamp, UV-transparent cuvettes, and a solution of purified DNA
39
absorbance to evaluate DNA purity
260 - 280 nm (based ni maam abi) 260 - most common
40
good quality DNA will have the ratio of?
1.7 - 2.0
41
ratio that indicates more contaminants present
<1.6
42
ratio indicates too much protein and is not acceptable
3
43
ratio indicates contaminants and still acceptable
2-2.5
44
strong absorbances around 230 nm can indicate that?
organic compounds or chaotropic salts are present
45
the __ the ratio, the ___ the amount of thiocyanate salt is present
the LOWER the ratio, the GREATER the amount of thiocyanate salt is present
46
absorbance best for greater than 1.5
260/320
47
a reading at **320nm** indicate?
turbidity in the solution and possible contamination
48
DNA concentration absotbance
260 nm
49
do not use ___ if no correction
A320

MolBio (8 decks)

Brainscape helps you reach your goals faster, through stronger study habits.
© 2025 Bold Learning Solutions. Terms and Conditions