Analysis Flashcards
(56 cards)
What does RFLP stand for?
Restriction fragment length Polymorphisms
What’s the difference between Northern and Southern blotting
Northern is uncut RNA
What does FISH stand for and what is it
Fluorescence in situ hybridisation is a modern method looking at gross changes to genes and chromosomes, uses a fluorescent or labelled probe hybridised to a specific target
What’s the main three steps in PCR
1) denaturation- heat 94degreesC for 2mins
2) annealing- cooling to 30-65 with excess of primers
3) DNA synthesis- addition of dNTPs catalysed by DNA polymerase
How can primers be detected?
Traditionally by staining of agarose with ethidium bromide or they can be labelled with:
NH2, CHO
Enzyme e.g horseradish, alkaline phosphatase, biotin, fluorophore
Name two thermostable DNA polymerases and their origins
1) taq polymerase from thermus aquaticus
2) pfu polymerase from pyrococcus furiosus
What does ASPCR stand for?
Allele specific PCR
What can be detected by multiplex PCR
Duchenne muscular dystrophy
What does RTPCR stand for
Reverse transcriptase PCR
What does real time PCR enable and what’s another name for it
Enables quantification of the product in real time also referred to as quantification PCR
What are 2 common short tandem repeats and what do they lead to
(CAG)n and (GCN)n leading to polyglutamine (Q) and polyalanine (A)
Name 3 natural processes that transfer DNA
1) transformation
2) transduction
3) conjugation
Name 3 mobile genetic elements
1) plasmids
2) insertion (IS) element
3) transposons
Define monocistronic
Translates single protein chain
Define polycistronic and what don’t make this type of DNA
Translates >2 protein chains which are organised in an operon, eukaryotes do not make polycistronic mRNA
Why can naturally occurring plasmids be helpful
1) resistance to antibiotics or toxic metals
2) metabolic functions
3) production of virulence factors
What do IS elements contain
Inverted repeat sequences plus a transposase
What are transposons
Similar to IS elements but contain additional functions (e.g antibiotic resistance genes)
Name a plasmid that contains multiple cloning sites (MCS) (restriction site)
pUC18/19
What’s the reporter gene in pUC18/19
LacZ used for blue & white screening
Name 4 other vectors
1) shuttle vectors
2) integration vectors
3) lambda cloning vectors
4) artificial chromosomes
What’s the problem with cloning eukaryotic DNA?
Bacteria can’t splice out introns
Define bioinformatics
In silico analysis of nucleotide and protein sequences
Give two types of DNA sequencing
1) maxam and Gilbert (chemical)
2) Sanger (enzymatic)
