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PatrickHeslip

EthanDanziger
SimoneSwetland

TestingPlantSubstanceasPotentialMedicines

Background:Plantsarealwaysbattlingforsurvivalinthewildespecially
againstdiseases.Duetothistheyhavedevelopedantimicrobialagentsto
defendagainstthemicrobes.Ifisolatedtheseantimicrobialagentscanbe
testedagainstmanyviruses.Thenifprovensafeforhumanscanbeused
tohelpkillthemicrobesthathaveinfectedthebody.

Purpose/Objective:Thepurposeofourprojectistotestwhethertheplant
willusedefensiveantimicrobials.Thenwewillseewhatspecific
antimicrobialsitwillusetofendoffthebacteriagrowth.

Materials:
Balance,weighboat,labscoops
LBbrothbase
Mediabottles,250mL
Sterilizer/autoclave
Waterbath,37,shaking
SterileLBagar
Laminarflowhoodanddisinfectant
Glasses,safety,plastic
Bunsenburnerandgaslighter
Inoculatingloop,Ni/Crwire
Petridishes,60X15mm,sterile
E.coli
JM109(stockplate)
Plantspecimen
Mortarandpestle
Pipet,10mLandpump
Plasticfunnelsshortstemmed

Filterpaperdisks,5mmdiameter
Beakers,100mL
Syringe,10mLandfilter,0.2um
Reactiontubesandrack,1.7mL
Methanol,absolute
Pipet,1mLandpump
Dryblockheater/heatblock
Forceps,finetipped
Ampicillin
Glassspreader
Incubatoroven,37
Lemonleaves

Procedure:
measure2gramsofplantmaterial
usingamortarandpestle,grindup2gramsofplanttissuewith10
millilitersofdeionizedwater
letsitfor3minutes
filterthesamplethroughan11centimeterfilterpaperfunnel
filtersterilizetheextractusingasyringefilter
collect1milliliterofextractina1.7millilitermicrotube.Labelthetube
Repeatthestepsusingmethanolinsteadofwater
FilterSterilization(procedure)
AttachprefittertosyringeandrinsewithH2O
Carefullyopensterilizationfilter,keepingthefilterinitsplastic
covering
Loadapprox1.5mLofH2Obasedfiltrateusingapipet
Depresstheplunger,Collectingthesterilefilteredfiltrateinto
microfugetube
Quicklyclosemicrofugecapuntilasnapsoundisheard
useflamesterilizationforcestotransferfiltersquaresintoH2O
controlandampicillincontrol
labelplate(bottom)

dispense1mLE.coliineachplate
Flamesterilespreadingloop
placefilterdiscsinplate

Results:

MethanolExtract:
24hrs.

48hrs.

WaterExtract:
24hrs.

48hrs.

24hours

48hours

MethanolExtract

Ampicillinpaperdisc
hasanantimicrobial
ringarounditandthe
plantextractsdont
havethering

Ampicillinpaperdisc
stillhasantimicrobial
ringarounditandthe
plantextractsare
startingtohave
antimicrobialring.
Therearewhitedots
onoursolution
becauseofcross
contamination.

WaterExtract

Ampicillinpaperdisc
hasanantimicrobial
ringarounditandthe
plantextractsdont
havethering

Ampicillinpaperdisc
stillhasantimicrobial
ringarounditandthe
plantextractsare
startingtohave
antimicrobialring.
Therearewhitedots
onoursolution
becauseofcross
contamination.

Analysis:

Ourmethanolextractandourplantextractbothendeduphavingan
antimicrobialringaroundit.Ourampicillinringalsohadanantimicrobial
ringaroundit.Wecouldtellbecausetherewasaclearcirclearoundthe
piecesofpaper.Unfortunatelyourtraysgotalittlecontaminatedduringthe
lab.Wehadwhitedotsonthebacteriafieldthatwasinthetray,butthat
didntaffecttheendresultsoftheproject.