}oaquin Canizales

Lab Paitneis:
uustavo Rios
Ninku Choi
Stephanye Alonso

Lab Section: 1227
Septembei 26, 2u1S



















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Aftei acquiiing all the uata, I piesenteu the absoibance measuiements my
inuiviuual gioup attaineu, anu plotteu it as a function of concentiation of methylene blue
(Figuie 1). The absoibance has a uiiect piopoitionality to the concentiation of methylene
blue. The uepenuent vaiiable in both giaphs is the Absoibance at a specific wavelength,
anu the inuepenuent vaiiable is the concentiation of methylene blue. Figuie 2 shows the
same ielationship as piesenteu in Figuie 1 since the two vaiiables aie the same. The
uepenuent vaiiable in Figuie 2 was the aveiage absoibance of each inuiviuual gioup fiom
the lab section, anu was then plotteu as a function of concentiation of the methylene blue
uye. Table 1 uemonstiates my peisonal uata, which helpeu ueteimine how much BNA I
extiacteu fiom a stiawbeiiy.




Figuie 1: A Stanuaiu Cuive of Absoibance veisus Concentiation of Nethylene Blue Bye.
Specific concentiation peicentages of methylene uye weie piepaieu; the absoibencies of
the mixtuies weie measuieu by a spectiophotometei at a wavelength of SSu nm. A tienu
line of y=11u.42x+u.uu87 with R
2
= u.96742 is fitteu.
u.uu
u.2u
u.4u
u.6u
u.8u
1.uu
1.2u
1.4u
u u.uu2 u.uu4 u.uu6 u.uu8 u.u1 u.u12
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Figuie 2: The Aveiage Absoibance of Nethylene Blue with Known Concentiations. Aftei
collecting all the uata within the lab section, the aveiage absoibance of methylene blue was
taken in iespect to the uye's coiiesponuing concentiation. A "best fit" tienu line of
y=117.64x+u.uS8S with R
2
= u.99748 was supeiimposeu.



Table 1: Amount of BNA Extiacteu fiom Stiawbeiiy. Aftei measuiing the absoibance of the
BNA mixtuie, the uata was useu to ueteimine the final yielu of BNA. The mean yielu of BNA
foi the lab section was u.u1SS-miciogiam g of stiawbeiiy with a stanuaiu ueviation of
+u.u1SS miciogiamg of stiawbeiiy.

We|ght of
Strawberry
(g)
Abs(SS0) for
Methy|ene
8|ue-sta|ned
DNA
so|ut|on
Concentrat|on
of So|ut|on ()
Concentrat|on
of DNA
(m|crograms]
mL)
1ota|
Amount of
DNA
kecovered
(m|crograms)
DNA |e|d
(m|crogram
DNA]g of
Strawberry)
31.0S 0.0S 0.00037 0.032 0.16 0.00S2


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D|scuss|on

1he relaLlonshlp expressed beLween Lhe absorbance and concenLraLlon from llgure 1 ls
dlrecLly proporLlonal. 1hls means LhaL by measurlng Lhe absorbance of a cerLaln mlxLure, Lhe
experlmenLer can deLermlne Lhe concenLraLlon of meLhylene blue, whlch can Lhen be
correlaLed Lo Lhe amounL of unA exLracLed. 1he reason why you can correlaLe Lhe
concenLraLlon of Lhe dye ln respecL Lo Lhe unA ls because Lhe exLracLed unA was placed lnLo a
soluLlon ln whlch Lhe meLhylene blue was added. upon Lhe addlLlon of Lhe dye, Lhe unA
molecules were bound Lo Lhe molecules of Lhe dye, Lherefore Lhe concenLraLlon of Lhe
meLhylene blue ls dlrecLly assoclaLed wlLh Lhe concenLraLlon of Lhe unA ln Lhe sample. Slnce
Lhe varlables are dlrecLly proporLlonal, Lhe slgnlflcance ls LhaL as one varlable lncreases or
decrease Lhe oLher varlable wlll maLch Lhe behavlor Lo keep Lhe relaLlonshlp consLanL. A
posslble source of error could have been LhaL my group dldn'L make Lhe mlx Lhe soluLlons
enough for Lhe concenLraLlon Lo be even LhroughouL Lhe cuveLLe. AnoLher source could have
come from Lhe specLrophoLomeLer my group used, as Lhe absorbance seemed Lo always be
changlng. 1he longer Lhe soluLlon was lefL lnslde Lhe machlne, Lhe more Lhe absorbance
seemed Lo change.
llgure 2 follows Lhe expecLed paLLern Lhe beLLer slmply because lL was an average of
absorbencles from Lhe lab secLlon. llgure 2 shows a greaLer value of 8
2
ln comparlson Lo llgure
1, whlch demonsLraLes LhaL Lhe Lrend llne of Lhe llgure 2 flLs beLLer wlLh Lhe daLa polnLs. 1hls ls
apparenL lf one compares boLh graphs, as Lhe daLa from Lhe second graph are closer Lo Lhe
Lrend llne Lhen Lhe one from Lhe flrsL graph, also Lhe besL-flL llne passes Lhrough Lhe error bars
LhaL only valldaLe Lhe facL LhaL Lhls seL of daLa follows Lhe expecLed paLLern. 1he error bars are
an obvlous characLerlsLlc of llgure 2, whlch represenL Lhe sLandard devlaLlon of each
absorbance ln response Lhe concenLraLlon of meLhylene blue. lor each of Lhe daLa polnLs Lhe
lndlvldual error bar shows how much Lhe lab secLlon devlaLed from Lhe mean value for LhaL
speclflc polnL. Cne posslble explanaLlon for Lhe devlaLlon of each polnL ls LhaL lndlvldual groups
dld or dld noL mlx Lhe soluLlons enough for Lhem Lo have conslsLenL concenLraLlons. AnoLher
source could have come from measurlng Lhe volume of each of Lhe concenLraLlon LhaL could
have led Lo a greaLer or lower value of concenLraLlon for each dlluLlon. 8oLh of Lhese facLors
could've caused a devlaLlon from Lhe expecLed values.
1he sLandard curve analysls proved Lo be very helpful for flgurlng ouL how much unA
was exLracLed from each sLrawberry. Slnce Lhe Lwo varlables where dlrecLly proporLlonal, Lhe
concenLraLlon of any mlxLure can be deLermlned by uslng Lhe equaLlon of Lhe Lrend llne. AfLer
deLermlnlng Lhe concenLraLlon, a relaLlonshlp beLween 0.001 of MeLhylene 8lue and 0.086
mlcrograms/mllllllLer of unA Lo converL Lhe concenLraLlon acqulred lnLo Lhe concenLraLlon of
unA. l can be sure LhaL whaL was measured was unA because Lhe dye LhaL was used blnds Lo
unA molecules. 8y removlng excess meLhylene blue and cleanlng lL wlLh Lap waLer, Lhe longer
unA sLrands wlll break down whlch allowed for Lhe measuremenL of unA concenLraLlon.
AlLhough Lhe amounL of unA l exLracLed was less Lhan Lhe mean, my number seems reasonable
because l dldn'L leL my unA lncubaLe ln Lhe dye for Lhe approprlaLe amounL of Llme. 1herefore
by noL allowlng Lhe unA Lo soak and blnd wlLh Lhe meLhylene blue Lhe absorbance measured
could have been slgnlflcanLly less causlng my resulLs Lo be less Lhan expecLed.
Conc|us|on
Slnce Lhe relaLlonshlp beLween Lhe Lwo varlables ls dlrecLly proporLlonal, lL was a loL
slmpler Lo deLermlne Lhe concenLraLlon of Lhe mlxLure. AfLer measurlng Lhe absorbance of Lhe
unA mlxLure l was able Lo quanLlfy Lhe amounL of unA LhaL was exLracLed. 1he varlaLlon for Lhe
class daLa was a blL large, buL l Lhlnk lL has Lo do wlLh Lhe many facLors LhaL could've caused an
error, such as Lhe slze of Lhe sLrawberry, Lhe quallLy ln whlch Lhe frulL was smashed, or any
measureable sLeps ln whlch Lhe lndlvldual sclenLlsL was aL faulL.
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