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SCGI Practicals
Isolation of Genomic DNA
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Created by
Priyani Lad
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Cards (8)
Cell
lysis
1.
Cell membrane
is
disrupted physically
or chemically
2.
Fluid
containing all the cell components including
DNA
is obtained
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Lysate
Fluid obtained after cell
lysis
containing all the cell
components
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Breaking
down cell components during cell lysis
1.
Lipids
are broken down by
detergents
and surfactants
2.
Proteins
are broken down by
protease
3.
RNA
is broken down using
RNase
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Concentrating
lysate
1. Lysate is treated with
concentrated salt solution
2. Broken components
clump
together
3. DNA freely
floats
in the solution
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Separating
DNA from debris
1. Solution is
centrifuged
2. Clumped
debris
is separated from
DNA
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DNA precipitation
1.
Ice cold alcohol
plus a salt is added to
increase ionic strength
2.
DNA precipitates
out
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DNA pellet
Pellet of DNA obtained after
centrifugation
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Suspending
DNA pellet
1. Pellet is suspended in slightly
alkaline
solution mostly TE buffer or
ultra-pure
water
2. For subsequent DNA experimentation usually
PCR
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