Draw bp-GLO bplasmid b- b bcorrect banswer- bGFP-->bla-->ori<---araC
function bof bbuffer bP1 b- b bcorrect banswer- blyses bbacterial bcells band breleases
bcytoplasmic bcontents binto bbuffer
function bof bbuffer bP2 b- b bcorrect banswer- bdenatures bchromosomal bDNA band
bproteins
function bof bbuffer bN3 b- b bcorrect banswer- badds bsalt bto bprecipitate bchromosomal
bDNA band bproteins
function bof bbuffer bPB b- b bcorrect banswer- ballows bDNA bto bbind bto bthe
bmembrane bof bthe bspin bcolumn
function bof bbuffer bPE b- b bcorrect banswer- bwash bbuffer bfor bcleanup bof bDNA
components bneeded bfor bPCR b- b bcorrect banswer- bEdvotek bPCR bpellet b
Small bPCR btube
Micropipettors band btips
Primers
Plasmid bDNA
Microcentrifuge
Thermal bcycler
10X bgel bloading bsolution
whats bin bthe bPCR bpellet b- b bcorrect banswer- bbuffer, bMg2+, bdNTPs, bTaq
bpolymerase
denaturing b- b bcorrect banswer- bThe bfirst bstep, b94 bdeg. bcauses bcomplementary
bbases bin bthe bdouble-stranded bDNA bhelix bto bcome bapart bresulting bin bsingle-
stranded bDNA bmolecules.
annealing b- b bcorrect banswer- bThe bsecond bstep, bat b58 bdeg. bthe bnucleotide
bbases bin bprimers banneal bto bthe bcomplementary bbases bpresent bin ba bDNA
btemplate.
extension b- b bcorrect banswer- bThe bthird bstep, bat b72 bdeg. bthe bTaq bDNA
bpolymerase bbinds bto bthe bend bof ba bprimer band bcopies bor b"extends" bthe
bopposite bstrand bof bDNA.
how bdoes bPCR bcreate bcopies b- b bcorrect banswer- bEvery bcycle bdoubles bthe
bDNA bsequence bof binterest
gel belectrophoresis b- b bcorrect banswer- banalyze bDNA bby bseparating bDNA
bmolecules baccording bto btheir bsize b(length bas bmeasured bin bbase bpairs).
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