[26] DNA and RNA Isolation Techniques Flashcards

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1
Q

What is DNA isolation?

A

The process of purifying DNA from a sample using a combination of physical and chemical methods.

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2
Q

What is RNA isolation?

A

The process of purifying RNA from a sample using a combination of physical and chemical methods.

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3
Q

What are the general steps in DNA isolation?

A
  • Cell lysis
  • Protein removal
  • DNA precipitation
  • DNA washing
  • DNA rehydration
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4
Q

What are the general steps in RNA isolation?

A
  • Cell lysis
  • Protein removal
  • RNA precipitation
  • RNA washing
  • RNA rehydration
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5
Q

What is cell lysis in the context of DNA and RNA isolation?

A

The breaking down of the cell membrane to release cellular contents.

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6
Q

Why is protein removal needed in DNA and RNA isolation?

A

Proteins can interfere with the analysis and processing of the DNA or RNA.

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7
Q

What is the purpose of DNA or RNA precipitation?

A

It allows the DNA or RNA to be separated from the rest of the cellular contents.

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8
Q

Why is washing necessary in DNA and RNA isolation?

A

To remove impurities and unwanted components.

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9
Q

What does rehydration of DNA or RNA mean?

A

It’s the step where the DNA or RNA is dissolved in a small volume of water or buffer for storage and further analysis.

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10
Q

What does the purity of DNA or RNA mean?

A

It refers to the degree to which the DNA or RNA is free from contaminants that may interfere with subsequent experiments.

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11
Q

What techniques are commonly used for DNA isolation?

A

Phenol-chloroform extraction, column-based extraction, magnetic bead-based extraction, etc.

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12
Q

What techniques are commonly used for RNA isolation?

A

Guanidinium thiocyanate-phenol-chloroform extraction, column-based extraction, magnetic bead-based extraction, etc.

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13
Q

What is phenol-chloroform extraction?

A

A method for purifying DNA or RNA, based on the differential solubility of DNA/RNA, proteins, and lipids in immiscible phases.

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14
Q

What is column-based extraction?

A

A method where DNA or RNA binds to the column, impurities are washed away, and the DNA or RNA is then eluted.

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15
Q

What is magnetic bead-based extraction?

A

A method where DNA or RNA binds to magnetic beads, impurities are washed away, and the DNA or RNA is then eluted.

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16
Q

What is guanidinium thiocyanate-phenol-chloroform extraction?

A

A method for RNA purification where the RNA remains in the aqueous phase, while DNA and proteins partition into the organic phase.

17
Q

Why is RNA isolation more difficult than DNA isolation?

A

RNA is more unstable and susceptible to degradation by enzymes called RNases.

18
Q

What is RNase?

A

An enzyme that degrades RNA.

19
Q

What precautions must be taken during RNA isolation?

A

To prevent RNA degradation, all tools and solutions must be RNase-free, and the process should be quick.

20
Q

What is DNA?

A

Deoxyribonucleic acid, the molecule that carries the genetic instructions for the development, functioning, growth, and reproduction of all known organisms.