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Q&A Why does the genomic
DNA band appear
smeared in agarose
gel electrophoresis?
Plasmid
DNA Extraction Kit(PD004, PD100, PD300) --Q&A
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Genomic
DNA Extraction Kit(GT004,GT050,GT300,GB004,GB100,GB300,GP004,GP100)--Q&A
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Why doesn¡¦t the
Eluted DNA perform
well in downstream
applications? |
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Why does the genomic
DNA band appear
smeared in agarose
gel electrophoresis? |
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Several points
should be noted to
avoid DNA
degradation:
(1) DNA degradation
occurs when the
sample is not fresh
or is stored
improperly for a
long time. Samples
not used immediately
should be flash
frozen in liquid
nitrogen and stored
at -80¢XC.
Genomic DNA in samples stored at room
temperature, 4¢XC, or
-20¢XC are subject to
degradation. It is
also not advised to
keep samples in
buffer or medium
while storing at
-80¢XC.
(2) For whole blood
samples, if they are
stored at room
temperature for more
than 2 days or at
4¢XC or -20¢XC,
isolated genomic DNA
appears smeared at
an extent
proportional to the
storage time.
(3) Use fresh TAE or
TBE running buffer
for electrophoresis.
Running buffer that
is used repeatedly
may be contaminated
with DNase.
(4) If isolated DNA
needs to be stored
for a long time, use
10 mM Tris-HCl (pH
9.0) or TE for
elution. Using ddH2O
is not advised in
this case as DNA
fragments in H2O
suffer from
gradual degradation through acid hydrolysis.
(5) If DNA is to be
used frequently,
elute in 10 mM
Tris-HCl (pH 9.0) or
TE and store at 4¢XC.
Keep DNA at -20¢XC
only for long-term
storage.
Repeated freeze-thaw cycles can cause shearing of
genomic DNA.
(6) Genomic DNA
extracted from
paraffin-embedded
tissue is usually
degraded. This is
because genomic DNA
in paraffin-embedded
tissue unavoidably
suffers from
degradation when
samples are treated and stored for a long time. DNA in
this case is not
suitable for
Southern blotting or
restriction analysis
due to smearing.
However, it is
applicable for PCR. |
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What is the key to
successfully
isolating genomic
DNA of good yield
and quality? |
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Why does the sample
appear viscous after
sample Lysis and not
pass through the
column easily, even
when less than the
suggested maximum
sample amount is
used? |
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Q |
When Buffy coat is
used, the column
tends to be clogged. |
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DNA Fragments Extraction Kit
(DF004, DF100, DF300)--Q&A
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