DNA Extraction and Analysis Methods

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36 Questions

What is the purpose of DNA extraction in medical genetics?

To identify mutations or diseases

What is the purpose of PCR in molecular biology?

To amplify a single copy or a few copies of a segment of DNA

What is the function of Taq DNA Polymerase in PCR?

Catalyses primer extension as nucleotides are added

What is the purpose of Tris-EDTA (TE) buffer in DNA extraction?

To prevent DNA from degradation

Which step of the PCR cycle involves denaturation by heat?

$1^{st}$ PCR Cycle - Step 1

What does PCR stand for?

Polymerase chain reaction

What is the main purpose of DNA isolation?

Purification of DNA from sample

What is the role of biotinylated primers in PCR?

Anneal to ends of target sequence

Which technique is used to make a large number of copies of a gene or gene segments?

Polymerase chain reaction (PCR)

What is the final product of PCR?

Thousands to millions of copies of a particular DNA sequence

What is the function of agarose gel in electrophoresis?

To provide a matrix for separating molecules like DNA and proteins

What is the role of ethidium bromide in DNA electrophoresis?

To intercalate between base pairs in DNA and emit fluorescence under UV light

What is the purpose of a DNA ladder in gel electrophoresis?

To determine the size of an unknown DNA sample by comparison with known sizes

What is the function of loading dye in gel electrophoresis?

To monitor the movement of the sample

What does a buffer solution contain in the context of gel electrophoresis?

A weak acid and its salt or a weak base and its salt, resistant to changes in pH

What is the primary factor that determines the migration of DNA in gel electrophoresis?

Molecule size

Electrophoresis separates molecules based on charge, not size or shape

False

Agarose gel is derived from animal sources

False

The DNA ladder is used to determine the charge of an unknown DNA sample

False

Ethidium bromide is used to detect DNA following gel electrophoresis

True

The power supply is a source of electric current for gel electrophoresis

True

DNA is positively charged and migrates towards the negative electrode in gel electrophoresis

False

The primary function of the loading dye is to prevent sample floating during electrophoresis

True

The micropipettes are used for transferring large volumes of reagents in molecular biology labs

False

DNA extraction involves only chemical methods and does not require any mechanical processes.

False

The purpose of PCR is to amplify a single copy or a few copies of a segment of RNA.

False

The function of Taq DNA Polymerase in PCR is to catalyze the extension of RNA primers.

False

The DNA storage buffer, Tris-EDTA (TE) buffer, is used in DNA extraction to prevent RNA degradation.

False

PCR Cycle - Step 2 involves denaturation by heat.

False

The purpose of DNA extraction is to isolate DNA for genotyping, forensics, and paternity testing, but it is not used in medical genetics to identify mutations or diseases.

False

PCR is a technique used in molecular biology to amplify DNA segments, but it cannot generate millions of copies of a particular DNA sequence.

False

What is the role of ethidium bromide in DNA electrophoresis?

It intercalates between base pairs in a double-stranded DNA molecule and emits fluorescence when exposed to UV light

What is the function of agarose gel in electrophoresis?

To separate molecules based on charge, size, and conformation

What is the primary factor that determines the migration of DNA in gel electrophoresis?

Molecule charge

What is the purpose of a DNA ladder in gel electrophoresis?

To determine the size of an unknown DNA sample by comparing it with known base pair sizes

What is the purpose of Tris-EDTA (TE) buffer in DNA extraction?

To prevent RNA degradation

Study Notes

  • DNA extraction: a process for purifying DNA from samples using physical and chemical methods, used in science and medicine for genotyping, forensics, paternity testing, and medical genetics to identify mutations or diseases.
  • Steps for DNA extraction: digestion (mechanical and chemical), precipitation with ice cold ethanol, and storage in Tris-EDTA (TE) buffer to prevent degradation.
  • PCR (polymerase chain reaction): molecular biology technique for amplifying a single or few copies of DNA segments, generating thousands to millions of copies for large-scale analysis.
  • PCR cycle: denaturation (heat) of target sequence, annealing of biotinylated primers to target ends, and primer extension by Taq DNA Polymerase with added nucleotides.
  • Exponential amplification: number of amplicon copies doubles with each cycle, e.g. 2 copies after 1 cycle, 4 after 2 cycles, 64 after 6 cycles.
  • DNA electrophoresis: procedure for separating DNA or proteins through a stationary gel in an electric field based on size, charge, and conformation.
  • Agarose gel electrophoresis: made from agar, a polysaccharide derived from seaweed, and ethidium bromide is used to detect DNA by staining and emitting fluorescence under UV light.
  • DNA ladder: a reference sample with known DNA sizes used to compare unknown DNA in gel electrophoresis, appearing as regularly spaced bands.
  • Precise pipetting: essential skill for molecular biology lab tasks, which involves accurately transferring small volumes of reagents using micropipettes.
  • Micropipettes: come in various sizes for aspirating small (1-5 uL), medium (20-50 uL), and large (100-500 uL) volumes.

Test your knowledge of DNA extraction, PCR, and DNA electrophoresis with this quiz. Learn about the process of purifying DNA from samples, its applications in genotyping, forensics, paternity testing, and medical genetics, as well as the steps involved in DNA extraction.

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