MCBG - Advanced Molecular Techniques Flashcards
Give some ways to analyse DNA at the nucleotide level.
- DNA sequencing
- PCR with restriction analysis
Give some ways to analyse DNA at the gene level.
- Southern and Northern blotting
- RT-PCR
- Microarray
- DNA fingerprinting/profiling
Give some ways to analyse DNA at the chromosome level.
- Karyotyping
- FISH/chromosome painting
What is DNA sequencing?
The process of determining the precise order of nucleotides within a DNA molecule.
Why is DNA sequencing an ethical issue?
- could open up areas for discrimination
- hard to decide who owns the sequence (person who sequenced it or patient themselves?)
What are allele-specific primers used for?
In PCR, allele specific primers can be used to create a PCR product which includes a certain mutation with a known location.
What is Southern Blotting used to show?
Presence of a particular DNA fragment in a sample.
What is the purpose of RT-PCR?
It is used to clone expressed genes by reverse transcribing the RNA into DNA through the use of reverse transcriptase.
How does DNA microarray work?
Samples of denatured DNA are labelled green (for individual) or red (control) with fluorescent dyes. They hybridise to a chip containing both normal and mutated ssDNA, and it can be used to show whether the individual has the mutation or not.
What are minisatellites?
Short sequences (10-60bp) of repetitive DNA that show variation from one person to the next.
What technique is used to amplify DNA so it can be profiled?
PCR
What is karyotyping?
A test to identify and evaluate size, shape and number of chromosomes in a sample of body cells.
What is FISH (fluorescent in situ hybridisation) used for?
To detect the presence/absence of specific DNA sequences on chromosomes.
