DNA manipulation Flashcards
3 stages of PCR
Denaturation, annealing, extenstion
what is required for PCR
taq polymerase, free nucleotides, primers, DNA
role of restriction enzymes/endonucleases
‘cut’ dna at a specific site
where do restriction enzymes often cut
at a 6 base palindrome
role of ligases
forms phosphodiester bonds between segments of DNA
where does taq polymerase come from
the bacteria thermus aquaticus
role of DNA polymerases
catalyse the addition of nucleotides to DNA strands
optimal temperature for taq polymerase
70-75º
temperature for denaturation
95º
temperature for annealing
around 50-70º
what is pcr
a technique used to amplify a particular segment of a DNA sample, and make a large amount of copies
what is gel electrophoresis
technique used to seperate DNA fragments based on their sizes
what charge does DNA have
negative (because of phosphate groups)
what is a DNA ladder
a sample of DNA fragments of known lengths, used to determine the lengths of other fragments in gel electrophoresis
what is a recombinant plasmid
a plasmid with DNA from the original organism, as well as DNA from another
