Bio-Rad Bio-Spin 6 and Micro Bio-Spin 6 Columns - Manual (Page 2)

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SIG 020996 Printed in USA
Buffer Exchange
The gel in the Micro Bio-Spin columns is suspended in either
SSC buffer, pH 7.0, or Tris buffer, pH 7.4. The gel matrix is
compatible with most aqueous buffers. Buffer exchange can be
achieved using the following procedure.
1. Follow steps 1 and 2 in the Instructions for Use section.
2. Apply the new buffer in 500 µl aliquots. After each appli-
cation of new buffer, let the buffer drain out by gravity, or
centrifuge the column for 1 minute to remove the buffer.
Discard buffer from collection tube. Repeat as required.
Three washes result in > 99% of the buffer exchanged. Four
washes result in > 99.9% of buffer exchanged.
3. Sample can now be applied to the column as directed in
steps 3 through 6 in the Instructions for Use section.
Centrifugation Notes
Micro Bio-Spin columns fit 1.5 ml microcentrifuge tubes for
sample collection during centrifugation. Use the 2.0 ml micro-
tubes provided with the columns for the initial column equili-
bration step.
Benchtop microcentrifuges capable of generating a minimum
force of 1,000 x (g) are suitable for Micro Bio-Spin column use.
The gravitational force created at a particular rpm is a function
of the radius of the microcentrifuge rotor. Consult the micro-
centrifuge instruction manual for conversion information from
rpm to g-force. Alternatively, to calculate the speed (rpm)
required to reach a gravitational force of 1,000 x (g), use the
following equation:
RCF (g) = (1.12 x 10 -5 )(rpm) 2 r
where r is the radius in centimeters measured from the center
of the rotor to the middle of the Micro Bio-Spin column, and
rpm is the speed of the rotor in revolutions per minute.
Sample Dilution Troubleshooting Guide
Sample dilution will occur if, during column equilibration, the
column is centrifuged without first removing the excess pack-
ing buffer. Dilution will also occur if the excess packing buffer
is left in the collection tube prior to centrifugation. The tip of
the column becomes submerged and retains excess buffer.
To correct this situation, always discard the buffer from the
collection tube after gravity draining and after the first cen-
trifugation (steps 1 and 2 of Instructions for Use section).
Sterilization
If a sterile Micro Bio-Spin column is required, autoclave the
column at 121 °C for 20–30 minutes. If exchanging buffers,
the buffer pH in the column should be in the range of 6.0 to
8.0 prior to autoclaving.
4006051 Rev B
Ordering Information
Catalog
Number
Product Description
Bio-Gel P-6 Columns in SSC
732-6200 Micro Bio-Spin 6 Column, SSC, 25, includes
25 P-6 columns in SSC buffer, 25 buffer col-
lection tubes, and instruction manual
732-6201 Micro Bio-Spin 6 Column, SSC, 100, includes
100 P-6 columns in SSC buffer, 100 buffer col-
lection tubes, and instruction manual
732-6205 Micro Bio-Spin 6 Column, SSC, 1,000,
includes 1,000 P-6 columns in SSC buffer, 1,000
buffer collection tubes, and instruction manual
Bio-Gel P-6 Columns in Tris
732-6221 Micro Bio-Spin 6 Column, Tris, 25, includes
25 P-6 columns in Tris buffer, 25 buffer collec-
tion tubes, and instruction manual
732-6222 Micro Bio-Spin 6 Column, Tris, 100, includes
100 P-6 columns in Tris buffer, 100 buffer col-
lection tubes, and instruction manual
732-6225 Micro Bio-Spin 6 Column, Tris, 1,000, includes
1,000 P-6 columns in Tris buffer, 1,000 buffer
collection tubes, and instruction manual
Bio-Gel P-30 Columns in SSC
732-6202 Micro Bio-Spin 30 Column, SSC, 25, includes
25 P-30 columns in SSC buffer, 25 buffer col-
lection tubes, and instruction manual
732-6203 Micro Bio-Spin 30 Column, SSC, 100,
includes 100 P-30 columns in SSC buffer, 100
buffer collection tubes, and instruction manual
732-6206 Micro Bio-Spin 30 Column, SSC, 1,000,
includes 1,000 P-30 columns in SSC buffer, 1,000
buffer collection tubes, and instruction manual
Bio-Gel P-30 Columns in Tris
732-6223 Micro Bio-Spin 30 Column, Tris, 25, includes
25 P-30 columns in Tris buffer, 25 buffer col-
lection tubes, and instruction manual
732-6224 Micro Bio-Spin 30 Column, Tris, 100,
includes 100 P-30 columns in Tris buffer, 100
buffer collection tubes, and instruction manual
732-6226 Micro Bio-Spin 30 Column, Tris, 1,000, includes
1,000 P-30 columns in Tris buffer, 1,000 buffer
collection tubes, and instruction manual
Empty Columns
732-6204 Empty Micro Bio-Spin Column, 100,
includes 100 empty columns, 100 buffer col-
lection tubes, and instruction manual