7.1 Flashcards
genome
total of all genetic material in an organism
pcr
polymerase chain reaction
amplification of dna in vitro (out of body)need
-Taq polymerase
duplicates DNA
-DNA
Original DNA to be replicated
-primers x2
a single strand of DNA that is complimentary to the target sequence allowing you to amplify any
-nucleotides
free nucleotides to make amplified DNA
how does pcr work
- use (DNA) primers / nucleotides
- use {high temperature / 95 ◦
C} for 30 seconds (to separate
DNA strands by breaking h bonds - use {lower temperature / 55 ◦
C} for 20 seconds (to bind
primers) - heat to 72°C with polymerase (to replicate DNA)
- repeat process (to obtain many copies of DNA)
heat tolrant DNA polymerase then replicates region of DNA
takes longer for polyerasition of nucleotides
dna sequencing
-The DNA is chopped into smaller pieces
-Double strands are separated into single strands
-PCR is used to amplify (replicate) the DNA.
-Terminator bases are added to the strands
-The coloured tags allow the bases to be read rapidly.
dna profiling
analysing DNA for variations
mostly identical
found in introns (non coding regions)
micro/mini satellites -short sequences which are repeated
—- identifacation of repeating patterns in the non coding introns of dna
sattelites
micro - 2-6 bases repeated 5-100x
mini - 10-100 bases repeated 50-many00x
same position but different number of repeats
gel electrophoresis
dna is cutby restriction enzyme
electrical current seperates dna pieces in gel
negative DNA is attracted to positive electrode
separated DNA are transferred to membrane
chemicals make the bands visible
gene probe
short DNA sequences labelled with a florescent molecule that are complimentary to specific DNA sequences
bind with DNA - hybridisation
southern blotting
alkine buffer solution added to gel
nylon filter paper placed over
DNA solution drawn to paper
leaving the solution as blots
DNA is denatured so strands separate
short tandem repeats
micro satalite regions used in dna identifcation
restriction endonuclease
cut dna to make short tandem repeats