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Q: Can I wash and
reuse the beads?
A: Yes you can!
Many protocols that recommend acid washing use concentrated hydrochloric acid.
This is fine for glass beads which are inert to hydrochloric acid, but it
reacts with the surface of zirconium oxides generating reactive ceramic
intermediates which may interfere with your experiment. This is why we STRONGLY
ADVISE AGAINST ACID WASHING ZIRCONIUM BASED BEADS. We also ADVISE AGAINST ACID
WASHING STAINLESS STEEL BEADS due to potential corrosion.
Q: How do I wash
the beads?
A: Washing in
standard laboratory detergent (like Alconox®), followed by rinsing with
deionized water to remove detergent is sufficient. Make sure that no "soapy"
bubbles remain before allowing the beads to dry.
Q: Are the beads
RNase free?
A: No. After you
wash them, you can make them RNase free by treating them with a reagent like
RNAZap® or RNaseAWAY®, followed by rinsing with nuclease free water. Allow the
beads to air dry in an oven or at ambient temperature.
Q: Will the beads
stick together when I wash them?
A: After washing,
the beads may stick together upon drying. Gently shake or move the container
with the dried beads until they flow freely.
Q: How many times
can I reuse the beads?
A: You may reuse
the beads many times. It is hard to say how many uses you will get due to the
variability in sample hardness, collisions with other beads, and running speed
in the Bullet Blender™. That said, you may use the beads until you find that
their appearance is not homogeneous or is unsatisfactory (i.e. pits in the beads
or discolorations). Typically, you can reuse the beads about ten times.
Q: Do the beads
need any preparation?
A: For protein or DNA applications, the beads
should be sterilized,
but do not require additional preparation.
However, when RNA extraction applications are
desired extra care must be taken to avoid
decomposition of RNA from exogenous RNase activity.
To ensure that beads are not the source of RNase
activity, we recommend washing the beads with RNase
Zap® (Ambion, Inc.) or RNase Away® (Molecular
BioProducts), then rinsing with nuclease free water,
followed by autoclaving.
Q: How do I
sterilize the beads?
A: They can be
autoclaved on dry cycle to sterilize them. They may stick, so shake the
container with the dried beads until they flow freely. Alternatively, they may be gamma
irradiated or exposed to ethylene oxide. Any manipulations of the beads after
sterilizing must use aseptic techniques in order to preserve the beads'
sterility.
Q: Which beads are
good for homogenizing which samples?
A: Check the
bead selection guide above. The two basic rules: (1) use beads of
approximately the same size as the size of your samples, to maximize the
collisions between beads and samples, and (2) use denser beads for tougher
samples.
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