Home
 Q&AWhy does the
obtained RNA
appear smeared
and degraded?
 Plasmid
DNA Extraction Kit(PD004, PD100, PD300) --Q&A
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RNA
Related
Products(RT050,RT100,RT300,RB050,RB100,RB300,RP050,RP100)--Q&A
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Q |
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|
Q |
Why does the
obtained RNA
appear smeared
and degraded?
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A |
Three critical
steps, if not
performed well can
cause RNA
degradation.
(1) Handling and
storing of samples.
(2) Disruption of
samples. (3) Storage
of eluted RNA.
(1) Most animal
tissues can be
processed fresh
(unfrozen). It is
important to keep
fresh tissue cold
and to process it
quickly (within 30
minutes) after
dissecting.
If samples cannot be processed immediately, it should
be flash frozen in
liquid nitrogen and
stored at -80°C.
Samples should be
handled with RNase-free
tools.
(2) When the sample
is disrupted,
disruption needs to
be fast and
thorough. Slow
disruption (e.g.
placing cells or
tissue in RB Buffer
without any
additional physical
shearing) may result
in
RNA degradation by endogenous RNase released
internally, yet
still inaccessible
to the protein
denaturant in RX
Buffer.
(3) After elution of
RNA with RNase-free
ddH2O provided in
the system, store
RNA at -80°C.
(4) Degradation of
RNA may also occur
during loading into
a gel. Use gel and
freshrunning buffer
prepared using DEPC-treated
ddH2O, as well as a
properly cleaned
geltray and
tank for electrophoresis. Adding EtBr directly into the
gel can also avoid
possibledegradation
of RNA that may
occur during gel
staining. |
|
Q |
Is total RNA
isolated by the
Total RNA Extraction
System free of
genomic DNA? |
|
Q |
What is the most
suitable kit for
Cytoplasmic RNA
extraction? |
|
Q |
What is the function
of the lysate filter
columns? |
|
Q |
Which kit is
suitable for RNA
extraction from
yeast? |
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