[26] DNA and RNA Isolation Techniques Flashcards
What is DNA isolation?
The process of purifying DNA from a sample using a combination of physical and chemical methods.
What is RNA isolation?
The process of purifying RNA from a sample using a combination of physical and chemical methods.
What are the general steps in DNA isolation?
- Cell lysis
- Protein removal
- DNA precipitation
- DNA washing
- DNA rehydration
What are the general steps in RNA isolation?
- Cell lysis
- Protein removal
- RNA precipitation
- RNA washing
- RNA rehydration
What is cell lysis in the context of DNA and RNA isolation?
The breaking down of the cell membrane to release cellular contents.
Why is protein removal needed in DNA and RNA isolation?
Proteins can interfere with the analysis and processing of the DNA or RNA.
What is the purpose of DNA or RNA precipitation?
It allows the DNA or RNA to be separated from the rest of the cellular contents.
Why is washing necessary in DNA and RNA isolation?
To remove impurities and unwanted components.
What does rehydration of DNA or RNA mean?
It’s the step where the DNA or RNA is dissolved in a small volume of water or buffer for storage and further analysis.
What does the purity of DNA or RNA mean?
It refers to the degree to which the DNA or RNA is free from contaminants that may interfere with subsequent experiments.
What techniques are commonly used for DNA isolation?
Phenol-chloroform extraction, column-based extraction, magnetic bead-based extraction, etc.
What techniques are commonly used for RNA isolation?
Guanidinium thiocyanate-phenol-chloroform extraction, column-based extraction, magnetic bead-based extraction, etc.
What is phenol-chloroform extraction?
A method for purifying DNA or RNA, based on the differential solubility of DNA/RNA, proteins, and lipids in immiscible phases.
What is column-based extraction?
A method where DNA or RNA binds to the column, impurities are washed away, and the DNA or RNA is then eluted.
What is magnetic bead-based extraction?
A method where DNA or RNA binds to magnetic beads, impurities are washed away, and the DNA or RNA is then eluted.
What is guanidinium thiocyanate-phenol-chloroform extraction?
A method for RNA purification where the RNA remains in the aqueous phase, while DNA and proteins partition into the organic phase.
Why is RNA isolation more difficult than DNA isolation?
RNA is more unstable and susceptible to degradation by enzymes called RNases.
What is RNase?
An enzyme that degrades RNA.
What precautions must be taken during RNA isolation?
To prevent RNA degradation, all tools and solutions must be RNase-free, and the process should be quick.
What is DNA?
Deoxyribonucleic acid, the molecule that carries the genetic instructions for the development, functioning, growth, and reproduction of all known organisms.
