Home
 Q&AWhy DNA recovery is
lower than expected?
 Plasmid
DNA Extraction Kit(PD004, PD100, PD300) --Q&A
Genomic
DNA Extraction Kit(GT004,GT050,GT300,GB004,GB100,GB300,GP004,GP100)--Q&A
Gel/PCR
DNA Fragments Extraction Kit
(DF004, DF100, DF300)--Q&A
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Q |
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Q |
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Q |
A band, smaller than
the desired DNA
fragment is present
in the eluant. |
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Q |
Why DNA recovery is
lower than expected? |
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A |
(1) Gel slice did
not dissolve
completely.
u Gel slice was too
big. If using more
than 300 mg of gel
slice, separate it
into multiple tubes.
u Raise temperature
of incubation to
60°C and extend
incubation time.
(2) Incorrect DNA
Elution Step.
u Ensure that
Elution Buffer was
added and absorbed
to the center of the
DF Column matrix
(3) Incomplete DNA
Elution.
u If the sizes of
DNA fragments are
larger than 10 kb,
use preheated
Elution Buffer
(60-70°C) during the
Elution Step to
improve the elution
efficiency.
(4) Do not overload
the column with too
much DNA. Higher
recovery is attained
when lower amount of
DNA is loaded.
Divide the large
amount of DNA into
more than one
column.
(5) If ddH2O is used
for elution, make
sure that its pH is
between 7.0 and 8.5,
as pH lower than 7
leads to lower
elution efficiency.
(6) Make sure that
complete DNA elution
takes place by
adding no less than
30 ml of elution
solution onto the
membrane and letting
it completely absorb
into the membrane
before
centrifugation.
(7) Large DNA
fragments are eluted
less readily than
small DNA fragments.
When the DNA product
is larger than 5 kb,
use the elution
solution preheated
to 60˚C. |
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Q |
Why doesn’t the
Eluted DNA perform
well in downstream
applications? |
RNA
Related
Products(RT050,RT100,RT300,RB050,RB100,RB300,RP050,RP100)--Q&A
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