Brainscape's Knowledge GenomeTM

Browse over 1 million classes created by top students, professors, publishers, and experts.


See full index

BIO107 - Cell and Molecular Biology > Lesson 3 - Nucleic Acid Isolation > Flashcards

Lesson 3 - Nucleic Acid Isolation Flashcards

You may prefer our related Brainscape-certified flashcards:
AP® Biology flashcards Biology 101 flashcards
1
Q

application of nucleic acid extraction

A
  1. DNA profiling
  2. molecular biotechnology
  3. phylogenetic studies
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
2
Q

where is high quality used for

A
  1. restriction digestion
  2. gene cloning
  3. amplification
  4. DNA sequencing
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
3
Q

Common sources of DNA

A
  1. whole blood
  2. hair
  3. sperm
  4. bones
  5. nails
  6. tissues
  7. saliva
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
4
Q

What are the five general steps in nucleic acid isolation

A
  1. tissue homogenization and cell lysis
  2. denaturation and separation of other biomolecules from nucleic acid
  3. precipitation of nucleic acid from aqueous phase
  4. washing of precipitated nucleic acid
  5. drying of pellet and dissolution of dried pellet
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
5
Q

mechanical method for tissue homogenization

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

refers to the process of applying sound energy to agitate particles or discontinuous fibers in a liquid

A

Sonication

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

reduce the size of materials to give a usable form or to separate their components

A

grinding

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

chemical used for cell lysis

A
  1. buffer
  2. salt
  3. detergent/surfactant
  4. denaturants
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
9
Q

buffer

A

Tris-HCl

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

salt

A

NaCl

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

detergent/surfactant

A

SDS (sodium dodecyl sulfate)

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

purpose of detergent

A

emulsify phospholipids

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

denaturants

A

guanidinium

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

what does guanidinium do

A

inactivates RNases / DNases

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

used to treat other cellular components

A

enzymatic treatment

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

enzymatic treatment examples

A
  • lysozyme
  • cellulase
  • pectinase
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
17
Q

chemical treatment for denaturation and separation of other biomolecules from the nucleic acid

A
  1. phenol
  2. choloroform
  3. isoamyl alcohol
  4. CTAB
  5. PVP
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
18
Q

denature proteins

A

phenol

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

what does phenol denature

A

proteins

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

removes proteins and lipids

A

chloroform

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
21
Q

what does chloroform remove

A

proteins and lipids

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
22
Q

removes phenol and chloroform

A

isoamyl alcohol

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
23
Q

what does isoamyl alcohol remove

A

phenol and chloroform

24
Q

removes polysaccharides

A

CTAB (cetyltrimethylammonium bromide)

25
Q

what does CTAB (cetyltrimethylammonium bromide) remove

A

polysaccharides

26
Q

removes polyphenols (alkaloids)

A

PVP (polyvinylpyrrolidone)

27
Q

what does PVP (polyvinylpyrrolidone) remove

A

polyphenols (alkaloids)

28
Q

used in enzymatic treatment during the denaturation and separation of other biomolecules from the nucleic acid

A
  • protease
  • proteinase
29
Q

sample will be separated by density with heavier stuff at the bottom of the tube and lighter stuff on top

A

centrifugation

30
Q

formed during centrifugation

A
  1. supernatant
  2. precipitate
31
Q

clear liquid free of precipitate located above the solid part

A

supernatant

32
Q

solid that forms at the bottom after centrifugation

A

precipitate

33
Q

monovalent cations

A
  • sodium
  • potassium
34
Q

alcohol used in the precipitation of nucleic acid from aqueous phase

A
  • ethanol (95%)
  • isopropanol (95%)
35
Q

used to wash precipitated nucleic acids

A

70% ethanol / isopropanol

36
Q

two types of dyring of pellet

A
  1. air drying
  2. vacuum drying
37
Q

where is dissolution of dried pellet done

A
  • sterilized molecular grade water or
  • TE (Tris-EDTA)
38
Q

EDTA

A

Ethylenediaminetetraacetic acid

39
Q

inactivates DNases

A

EDTA

40
Q

what does EDTA inactivate

A

DNases

41
Q

after isolation,
__ treatment if DNA is isolated

A

RNA

42
Q

after isolation,
__ treatment if RNA is isolated

A

DNA

43
Q

storage of isolated DNA

A
  1. stock solution
  2. working solution
44
Q

stock solution temp.

A

-20°C to -80°C

45
Q

working solution temp.

A

4°C

46
Q

stock solution

A

pellet + Tris-EDTA

47
Q

how to determine purity and concentration of DNA isolation

A
  1. using DNA standard (different concentrations)
  2. UV spectrophotometer (purity)
48
Q

laboratory method used to separate mixtures of DNA, RNA, or proteins according to molecular size.

A

Gel electrophoresis

49
Q

To assess purity using the UV spectrophotometer, what is done?

A
  1. take readings at 260 nm - 280nm
  2. get ratio of A260/A280
  3. get ratio of A260/A230
50
Q

high purity for NA sample

A

A260/A280 = 1.8 to 2.0

51
Q

A260/A280 < 1.8 (or <1.6)

A

protein contamination

52
Q

protein contamination

A

A260/A280 < 1.8 (or <1.6)

53
Q

A260/A280 > 2.0

A

chloroform/phenol contamination

54
Q

chloroform/phenol contamination

A

A260/A280 > 2.0

55
Q

if A260/A230 < 2.0, it is contaminated with:

A
  1. carbohydrate carryover
  2. residual phenol/guanidinium
  3. other organic compounds
  4. salts

BIO107 - Cell and Molecular Biology (13 decks)

Brainscape helps you reach your goals faster, through stronger study habits.
© 2024 Bold Learning Solutions. Terms and Conditions