DNA probes Flashcards
What is a DNA probe?
a short single strand of DNA labelled with either a radioactive or fluorescent marker, with bases complementary with required gene
What reaction is used to make DNA probes?
PCR
Describe the DNA sequence that is needed to make a DNA probe
short, with complementary base sequence to the desired allele
Why is it important to wash away unattached DNA probes?
so unbound DNA probes are washed away, therefore there is not a false positive result
How would you determine the base sequence for the mutant allele (the allele that causes the disease)?
DNA sequencing/DNA library
What is DNA hybridisation?
when a section of DNA or RNA binds with a single stranded section of DNA with complementary bases.
Describe how radioactive probes could be used to show that a gene is present within a fragment of DNA?
- expose bases (form single strands)
- probe will bind to gene/sequence of alleles
- use autoradiography to show bound probes
How might a complementary strand of mRNA affect the production of a protein by a gene?
- complementary bases would form hydrogen bonds
- ribosome could not bind, preventing translation
How is DNA broken down into smaller fragments?
using restriction endonucleases, which cuts DNA at a specific point in the base sequence
Why might identifying the strain of bacteria causing a disease help improve treatment?
- can determine if resistant to antibiotic, so treatment will be effective
How could cDNA stop the production of a protein?
- will bind to mRNA
- forming double stranded mRNA
- this will prevent the ribosome from joining
- preventing translation
Why are probes needed?
because DNA is invisible on a membrane
Why are exons used, not introns?
- introns are not translated
- mutations in exons effect a/a sequence and therefore the 3° structure of a protein
- important to know whether or not an exam is effective rather than an intron
How can DNA be used to scan for harmful mutations?
- amplify using PCR
- cut using RE
- separate using gel electrophoresis
- add DNA probes
- show under UV or X-ray films