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ZYMO RESEARCH CORP.
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Lysis Methods
•
When using a Disruptor Genie
â
, vortex adapter, vortex, or
similar processing times will vary. Suggested beat beating time
is anywhere from 5-20 minutes. Calibrate bead beating times
to your particular device and application by testing several
different time points before using precious samples.
(Suggested times to test: 10, 20, and 30 minutes.) See
Appendix C for details.
•
When using high-speed bead bashing devices (e.g. Bertin
Percellys, MP – FastPrep24) run max speed for 5 minutes to
ensure unbiased lysis.
Incomplete Debris Removal
•
For high density samples, ensure lysate is centrifuged properly
to pellet insoluble debris following bead beating. Ensure that
none of the debris is transferred to the
Zymo-Spin™ III-F
Filter
in the next step.
Low DNA Yield
Input
•
If the lysate does not pass through the column or is extremely
viscous, use less input material. Too much sample input can
cause cellular debris to overload the column and insufficient
flow
•
Consult the Sample Input Table on Page 4 for information on
your particular input limit based on sample.
Binding Step
•
Ensure that the
ZymoBIOMICS™ DNA Binding Buffer
is
completely mixed with lysate before loading onto the column.
Improperly mixed samples can lead to poor DNA recovery.
Elution Procedure
•
Ensure the
ZymoBIOMICS
™
DNase/RNase Free Water
hydrates the matrix for at least 1 minute before centrifugation.
•
To increase yields, heat the
ZymoBIOMICS
™
DNase/RNase
Free Water
to 60ºC before use. Additionally, users can reload
the eluate onto the column matrix, incubate at room
temperature for 3 minutes, and centrifuge again to increase
yield without further dilution.
