GCU MICROBIOLOGY LAB FINAL
QUESTIONS WITH ANSWERS
competency b- b bcorrect banswer- bphysical bstate bthat ballows bDNA bto benter bthe
bcell
Describe bthe btransformation bpreformed bin blab b- b bcorrect banswer- b1. bLabel btest
btube b+pGLO band banother b-pGLO band bplace bthem bin bthe bfoam btube brack.
2. busing bsterile btransfer bpipettes, btransfer b250 buL bof btransformation bsolution
b(CaCl2).
3. bPlace bthe btubes bon bice.
4. bUse ba bsterile bloop bto bpick bout ba bsingle bcolony bof bbacteria bfrom bthe bstarter
bplate band btransfer bit binto bthe b+pGLO btube. b
5. bPlace bthe btube brack bin bthe bice. bUsing ba bnew bsterile bloop, brepeat bthe bstep
bfor b-pGLO btube.
6. bTransfer ba bsterile bloopful bof bpGLO bplasmid bDNA binto b+pGLO btube. bMix bwell
band breturn bthe btube bto bice.
7. bIncubate btubes bin bice bfor b10 bminutes.
8. bTransfer btubes bto bwater bbath bset bat b42o bC band bincubate bfor b50 bseconds.
bTransfer btubes bback bto bice band bincubate bfor btwo bminutes.
9. bAdd b250 buL bof bLB bnutrient bbroth bto bthe btubes. bIncubate bfor b10 bminutes bat
broom btemperature.
10. bTransfer b100 buL bof btube bcontents bto bappropriately blabeled bplates:
+pGLO bLB+amp bplate
+pGLO bLB+amp+ara bplate
-pGLO bLB+amp bplate
-pGLO bLB bplate
11. bSpread bthe bcontents bevenly bover bthe bsurface bof bagar. bInvert bplates band
bincubate bat b37o bC bfor b24 bhours.
12. bPlates bare bviewed bin bthe bdark bunder bUV blamp.
, Characteristics bof bplasmid b- b bcorrect banswer- bsmall bcircle bthat bcarries bthe
bGreen bFluorescent bProtein bgene b(GFP), bthe barabinose brepressor bgene b(araC),
band bthe bbeta-lactamase bgene b(bla).
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).
QUESTIONS WITH ANSWERS
competency b- b bcorrect banswer- bphysical bstate bthat ballows bDNA bto benter bthe
bcell
Describe bthe btransformation bpreformed bin blab b- b bcorrect banswer- b1. bLabel btest
btube b+pGLO band banother b-pGLO band bplace bthem bin bthe bfoam btube brack.
2. busing bsterile btransfer bpipettes, btransfer b250 buL bof btransformation bsolution
b(CaCl2).
3. bPlace bthe btubes bon bice.
4. bUse ba bsterile bloop bto bpick bout ba bsingle bcolony bof bbacteria bfrom bthe bstarter
bplate band btransfer bit binto bthe b+pGLO btube. b
5. bPlace bthe btube brack bin bthe bice. bUsing ba bnew bsterile bloop, brepeat bthe bstep
bfor b-pGLO btube.
6. bTransfer ba bsterile bloopful bof bpGLO bplasmid bDNA binto b+pGLO btube. bMix bwell
band breturn bthe btube bto bice.
7. bIncubate btubes bin bice bfor b10 bminutes.
8. bTransfer btubes bto bwater bbath bset bat b42o bC band bincubate bfor b50 bseconds.
bTransfer btubes bback bto bice band bincubate bfor btwo bminutes.
9. bAdd b250 buL bof bLB bnutrient bbroth bto bthe btubes. bIncubate bfor b10 bminutes bat
broom btemperature.
10. bTransfer b100 buL bof btube bcontents bto bappropriately blabeled bplates:
+pGLO bLB+amp bplate
+pGLO bLB+amp+ara bplate
-pGLO bLB+amp bplate
-pGLO bLB bplate
11. bSpread bthe bcontents bevenly bover bthe bsurface bof bagar. bInvert bplates band
bincubate bat b37o bC bfor b24 bhours.
12. bPlates bare bviewed bin bthe bdark bunder bUV blamp.
, Characteristics bof bplasmid b- b bcorrect banswer- bsmall bcircle bthat bcarries bthe
bGreen bFluorescent bProtein bgene b(GFP), bthe barabinose brepressor bgene b(araC),
band bthe bbeta-lactamase bgene b(bla).
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).
