genetic engineering 3 Flashcards
what does PCR stand for?
polymerase chain reaction
what is PCR?
method used to amplify minute amounts of DNA by repeated cycles of invitro replication
WHAT CAN pcr be used for?
creating mutations, detecting and amplifying minnute amounts of dna
what is required for PCR?
template(dna to be copied), sequence and start and end of DNA (2 primers), DNA polymerase, free nucleotides + termocycler
give 3 steps of PCR
denaturisation of DNA - strands separate
annealing of primers to dna
elongation of primers to template
give temperatures of PCR
95
55
72
how can PCR induce mutations?
primers can incorperate into DNA and insert sequence mutation
primers can add restriction enzyme points.
what is vecotr?
dna molecule that is maintained + replicated naturally by
organism (e.g.bacterial plasmid)
(can be usd to carry mtautions into organsim)
what is insert?
dna of interest to be cloned
give 3 essential properties of a cloning plasmid
have to contain origin of dna replication,
have restiction sites
have antibiotic restistance gene
what is frst step of cloning using plasmids ?
generation of campatible ends in insert and plasmid by restriction enzymes
what is enzyme that fuses fragment and plasmid?
dna ligase
what is tranformation?
where genetoc material is altered by incorperation of foreign dna into genome
what are 2 typesof plasmid?
cloning plasmid and expression plasmid
what is difference between bacterial and mammalian expression plasmids?
DNA expression plasmid has bacterial promoters
mammalian expression plasmid have eukaryotic promoters
