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MOLECULAR BIOLOGY - Lab > DNA extraction And Measurement > Flashcards

DNA extraction And Measurement Flashcards

1
Q

Uses of DNA extraction

A

Forensic
Medical
Scientific research

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2
Q

Basic steps in DNA extraction

A

Cell lysis
DNA precipitation
DNA purification

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3
Q

Mechanical methods of cell lysis

A

Grinding
Bead beating
Shearing
High pressure

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4
Q

Different chemical treatments

A

Buffer
Salt
Sodium Dodecyl Sulfate (SDS)
Senaturants
Reducing agents

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5
Q

Enzymatic treatments for cell lysis and what they are used for:

A

Proteinase K - animal cells
Cellulase - plant cells
Lyticase - yeasts
Lysozyme - bacteria

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6
Q

Methods of precipitating nucleic acids:

A

Monovalent cations
Alcohol precipitates
Centrifugation

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7
Q

What is a monovalent cation? Give examples

A

It neutralizes the (-) charged phosphate groups

Ammonium, potassium, sodium acetate, or lithium or sodium bromide

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8
Q

Ideal absorbance at A260/A280 ratio

A

1.8-2.0

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9
Q

What are the two types of spectrophotometer?

A

Traditional
NanoDrop

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10
Q

Maximal absorbance for DNA and RNA

A

260 nm

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11
Q

Maximal absorbance for proteins

A

280 nm

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12
Q

A highly efficient spectrophotometer that is able to accurately read the concentration and purity of the sample

A

NanoDrop spectrophotometer

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13
Q

Sample volume needed for NanoDrop spectrophotometers

A

0.5–2 uL

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14
Q

At what absorbance ratio is salt tested?

A

A260/A230

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15
Q

In wavelength indicators, what is the term used for referring the lower point on the spectrum?

A

Wavelength trough

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16
Q

In wavelength indicators, what is the term used for referring the highest point on the spectrum?

A

Peak

17
Q

Nucleic acids are resuspended in what?

A

Distilled water or Tris-EDTA

18
Q

Particles that forms when detergent molecules aggregate in a solution

A

Micelles

19
Q

What is the role of B–mercaptoethanol?

A

Reducing agent. Breaks sulfide bonds

20
Q

Volume ratioof phenol: chloroform: isoamyl alcohol

A

25: 24: 1

21
Q

Contaminants are denatured and accumukated in what phase?

A

Organic phase

22
Q

Nucleic acids are preserved in _____ phase

A

Aqueous

23
Q

What are the standard alcohols used for nucleic acid extractions?

A

Ethanol
Isopropanol

24
Q

What are the quantification methods of nucleic acids?

A

Spectrophotometric
Agarose gel electrophoresis

MOLECULAR BIOLOGY - Lab (1 decks)

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