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OPS Diagnostics, LLC
291 Route 22 East
Salem Industrial Park
Building 6
Lebanon, NJ 08833
TEL: (908) 253-3444
FAX: (908) 575-1660
Email:
info@opsdiagnostics.com
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Application Notes >Application
Table
There are many ways to prepare soy beans for protein extraction, DNA, and
RNA isolation. Here are several protocols using high throughput homogenizers
and the CryoGrinder.
References
Comparison of Methods for DNA Extraction from Soy Beans. OPS
Diagnostics Application Notes
Mixer Mill Preparation
Mixer mills such as the
Geno/Grinder and
HT Homogenizer are used when processing
a large number of samples is
necessary. Soy beans plants can be homogenized using bead beating in
several ways: dry, wet, or rehydrated. Rehydrating is a simple process
where by dry beans are soaked in a distilled water bath for 12 hours
prior to homogenizing. The next steps in the mixer mill preparation
process can be used with all three formats.
1. Place the bean in a 4 ml vial. Add either a 3/8" stainless steel or
tungsten carbide ball to the vial. Then cap the vial and place back
into a rack like a vial set.
2. Add about 500ul of buffer solution if homogenizing a rehydrated or
wet soy bean.
3. Place the rack into the mixer mill and process on high for 5 minutes
(Geno/Grinder or HT Homogenizer).
4. Balls can be removed from the Vial Set using the
24 Pin Magnet Ball
Dropper.
5. Remove the supernatant with the appropriate sized pipette and
transfer to a new vessel if desired.
6. Up to 15 dry soy beans can be processed using a 15ml vial sets.
These larger vials make use of two 7/16" stainless steel grinding balls.
7. Take note: that mixer mill disruption does generated heat and may
cause denaturing of certain enzymes and protein. Heat can be mitigated
by submersing the tubes and cryogenic rack into liquid nitrogen before
homogenization. Cryoblocks are available and act as cold sinks for
cryogenic grinding.
Grinding with CryoGrinder (miniature mortar and pestle)
Pulverization of leaf punch sample size
proportion can be achieved using the CryoGrinder. This method is best
used when high sample yield is less important. It can be used in
combination with liquid nitrogen.
1. Dry soybeans can simply be placed into the mortar and ground to a
fine powder using a small pounding method and then a circular motion
with the pestle.
2. Wet or rehydrated soybeans are much easier to grind. They can be
ground with less effort using the same technique listed above. In
addition other parts of the soy plant can be ground in a similar
fashion.
3. The mortar, pestle, and sample can be chilled if enzyme, DNA, or RNA
extraction is the main objective of soy disruption. Pre-chill the
mortar and pestle before adding the sample. Next, the sample can either
be chilled in a separate container or in the mortar itself. Take the
necessary safety precautions when handling the liquid nitrogen. Wearing
gloves, eye protection, and lab coat are highly recommended.
96 Deep Well Plates
Homogenization of leaf punch size
samples can also be done in 96 well format using a 96 well plate and
5/32"ss grinding balls.
1. Place the leaf punch or equal size sample into the well.
2. Then add 100ul of buffer solution
3. Next add the grinding balls using the 5/32"ss Ball Dropper.
4. Once homogenization has finished remove the balls by using a Magnet
Tip and recover the supernatant with the appropriate volume pipette.
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