detecting mutation Flashcards
Name the ways in which mutations can be detected
PCR Gel electrophoresis RFLP ARMS DNA sequencing
What is the point of PCR?
Allows amplification of DNA
Used for DNA cloning, DNA sequencing, gene identification and forensic medicine
What are the advantages of PCR?
Fast
Easy
Sensitive
Robust
What are the steps in PCR
It has three stages denaturation, anneal and extend
What is required for PCR?
Sequence information Oligonucleotide primers, DNA, Nucleotides and DNA polymerase
What does gel electrophoresis allow you to do?
Separation of DNA
What does restriction fragment length polymorphism allow you do to?
Identify variation in DNA by using restriction endonucleases to recognisee and cut DNA
Why is RFLP good?
Simple
Cheap
Why is RFLP bad?
Requires gel electrophoresis
What does ARMS allow you to do?
Detect mutations
How does ARMS work?
A special primer is added that will replicate a chain of DNA if the wild type (normal) mutation is present and another special primer is added that will replicate a chain of DNA if the mutation is present. The presence of absence of these replicated chains can be used to determine if the mutated gene is present.
Why is ARMS good?
cheap
no labelling is required
Why is ARMS bad?
Require sequence information and there is a limited amplification size.
What is DNA sequencing used for?
Detecting mutations using dideoxy nucleotide’s
Why is DNA sequencing bad?
Expensive equipment
