Basic Nucleic Acid Extraction Methods Flashcards

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1
Q

Beer-Lambert’s Law

A

A = log¹⁰ (I⁰/I) = ecl

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2
Q

Steps in DNA extraction

A
  1. Pretreatment and washing
  2. Cell lysis
  3. Removal of contaminants
  4. DNA precipitation
  5. DNA suspension
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3
Q

Physical methods on cell lysis:

A

Grinding
Shearing
Bead beating
Freeze-thaw

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4
Q

Types of solution-based method on cell lysis

A

Chemical
Enzymatic

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5
Q

What does SDS mean, and how does it remove contaminants in DNA extraction?

A

Sodium dodecyl sulfate

It makes the proteins anionic

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6
Q

A broad-spectrum serine protease that can also degrade nucleases

A

Proteinase K

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7
Q

What is used to remove RNA from a DNA extraction?

A

RNAse

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8
Q

Extraction methods on DNA precipitation

A

Organic
Inorganic
Solid phase

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9
Q

How does the organic method separate DNA from other components in a solution?

A

Uses high salt concentration in low pH with 1:1 phenol-chloroform

Solvation shell forms around the molecules

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10
Q

Inorganic method is also called as ______

A

Salting out

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11
Q

Optimal ratio of A260/A230 for DNA & RNA

A

2.0-2.2

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12
Q

Optimal ratio of A260/A280 for DNA & RNA

A

1.8-2.0

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13
Q

When dissociating a tissue, the sample must be _______ to inactivate intracellular RNAses

A

Frozen in liquid nitrogen or immeresed in a buffer solution

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