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DNA by DPA Method

1) The document describes an experiment to estimate the amount of DNA present in pea samples using the DPA (diphenylamine) method. 2) DNA is extracted from pea samples and different volumes are reacted with DPA reagent. A standard curve is prepared using known concentrations of DNA. 3) By comparing the absorbance of pea sample reactions to the standard curve, the concentration of DNA present in the pea samples is estimated to be 240 micrograms per ml.

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Shree Krishna Tarai
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251 views

DNA by DPA Method

1) The document describes an experiment to estimate the amount of DNA present in pea samples using the DPA (diphenylamine) method. 2) DNA is extracted from pea samples and different volumes are reacted with DPA reagent. A standard curve is prepared using known concentrations of DNA. 3) By comparing the absorbance of pea sample reactions to the standard curve, the concentration of DNA present in the pea samples is estimated to be 240 micrograms per ml.

Uploaded by

Shree Krishna Tarai
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PDF, TXT or read online on Scribd
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Page No.

CsnMATION OfOF DNA


DNA Experiment No. :......
BY DPA
AlM : To
tmETHOD
eshimae the Oumount
aumount of
qiven
samples c peas DA present
present in tbe
Liver by DPA me ttod
INTRODetION:
DNA s the
cels o Qnimals
qenehc mateial present in te
plants, prokaayoes and a feo
vimuses. Tn lukayotes it is ofen tound
voith prpeins to fom nucleo osDiattd
-prolins . In prokalyott s
the qenehc
qene hc matual has a single
DrdA Obout laiqe mole cule of
loDo
microme es in lenqth oith a nmbe
of Rarti proleins.DNA is most present in chomoso!
Contained withio he no cleus in Evkasyoks but it is also
Secn in
cutoplas mic orqanells Bukasycks.
such as chloroplast ancd
mitoch ond na but foy a few viuses Buch as backesicphagc
DNA js clouble skanded.

PRINCIPLE
DNA is xiaced usinq TCA. The amount of DNA
extsacted can be eshmated by DPA melhod (Diphenyl
amine nmetod) . To tais method the suqar compon ent
presence of an
de oryn bose rs prrseot in DNA in the
hu aldebyde
Qciel fomg huclroxgl Revulinil alde hyde.
acid a blue to
eacs diphun
with amine
Teageot mea sused
lbut cemplex. The blue
toloul colourfoam is
a suitable blank.
by reading at 600nm geainst
Pooja
NO

Re&UIREMENTS :

MieropipeMes andl tips


Boiling wake bath
2

Srond asd oNA soluhoo:


100 mg of DNA is diss olved io

OPA Teagnt
lqm DPA is digolved io to0ml of
To thi's add 0.7ml o Conc. , acehc aad
SO4
s | qm of Peas
Pestel morta cenkifuqe tubes, testhbes, pipeles
aod colonmetes.

PRocepuRe :

5TEP-1: Prpasahon of atandasd qraph.


Oifeent alligucks of standasd DrA tolwhon (ioc0 419 /n)
Tanqlng forom o2- Iml plpetted out into fve
The volune in eacb
dfeent testtubes in duplicates. oakes.
Mese tstubes is made upto Im) toith dishlled
added to all t e teshubes
4ml ef DPA Teaqent
bated
was
for lo minutes io boiled
Ihee testubes wes incu
The teshebes weie
walerbath for colour development
Cooled and Was read at e00nm
and concenhahon
Hed tati ng
Of DNA Io
in -axis.

Poojd
Page No. : .....2. Experiment No. :

STer -2: eshimation of oNA in Peas

Ol m of 9ample peas exhact uoese tokeo in oo


different testtubes and he volumne wese made Luptowas
Iml wsinq dishlled cate. 4m) of DPA reagent
added to tothe testtube and incu batedl tor 10minutes
waterbath. lhe tubes uoese cooled and Dwas
in
Frpm
read at 600nm aqainst a suitable blank.
DNA for bolh he sa mples
standasd graph te amount of
weu es hinatedl

ResuL:
wntnoon
The amount DaA pretnt
Somple is foung to be 240 Aiq lm
Ho
DNA
K Nucleo
Laeg HaPo

8la coudaucaki DPA

CH CooH
oH
DAA lau

-Ho

6
Dbservah'on toble

vol
TT No. vol o std vol of
Daly( std DNA DPA Cm) Absoba
DNA ( (salo
0-0 0g-Tncubae 00
in boik'ng
0-8 200owalc 36
2ampdetedl balß
3
400for
4 ml10 minob
0.69

4 b.6 D 4 600 b)

30
6
0 1000
1.60
1.0 Cuk)
0-42

NH
NH

NA Blue colcusad
Cole
::
Date T

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