Bio-Rad Bio-Spin 6 and Micro Bio-Spin 6 Columns - Manual

Introduction
Micro Bio-Spin chromatography columns are ready to use for
rapid and efficient cleanup and purification of nucleic acids
and proteins using a microcentrifuge.
Micro Bio-Spin Columns
• Remove dye terminators
• Remove unincorporated nucleotides
• Desalt nucleic acids, proteins and peptides
The columns are packed with special grades of Bio-Gel ® P poly-
acrylamide P-6 or P-30 gel matrices manufactured specifically
for Bio-Rad spin columns. This unique gel produces very effi-
cient, non-interactive size separations. Micro Bio-Spin columns
are suitable for use with 1.5 or 2.0 ml microcentrifuge tubes and
are completely autoclavable.
Technical Information
Gel Matrix
Bio-Gel P-6 or P-30 polyacrylamide gel suspended in 1.0 ml
of buffer
Buffers
SSC buffer (150 mM sodium chloride, 17.5 mM sodium cit-
rate, pH 7.0) with 0.02% sodium azide
Tris buffer (10 mM Tris-HCl, pH 7.4) with 0.02% sodium
azide
Sample Application Volumes
Nucleic acids, proteins, and peptides, 20–75 µl. Volumes less
than 20 µl may affect recovery
Exclusion Limits
Bio-Gel P-6 gel: 5 base pairs (nucleic acids) or 6,000 daltons
(proteins,peptides)
Bio-Gel P-30 gel: 20 base pairs (nucleic acids) or 40,000 dal-
tons (proteins, peptides)
Expected Retention and Recovery
Micro Bio-Spin 6 column
98% retention of unincorporated nucleotides at 20 µl
90% recovery of applied DNA at 20 µl
Micro Bio-Spin 30 column
100% retention of unincorporated nucleotides at 20 µl
95% recovery of applied DNA at 70 µl
Centrifuge Type
Microcentrifuge with a centrifugal force of 1,000 x (g).
Autoclavability
Micro Bio-Spin columns, Bio-Gel P gel, and collection tubes are
completely autoclavable at 121 °C for 30 minutes at pH 6.0–8.0.
Chemical Stability
pH 2–10, common aqueous buffers, formamide, dilute organic
acids, alcohol, 20% (V/V) other chaotropic agents, detergents.
Storage
Store at 4 °C. Do not freeze. Shelf life is 1 year at 4 °C.
Instructions for Use
1. Invert the column sharply several times to resuspend the
settled gel and remove any bubbles. Snap off the tip and
place the column in a 2.0 ml microcentrifuge tube
(included). Now remove the top cap. If the column does
not begin to flow, push the cap back on the column and
then remove it again to start the flow. Allow the excess
packing buffer to drain by gravity to the top of the gel bed
(about two minutes). Discard the drained buffer then
place the column back into the 2.0 ml tube.
2. Centrifuge for 2 minutes in a microcentrifuge at 1,000 x (g)
(see Centrifugation Notes section) to remove the remain-
ing packing buffer. Discard the buffer.
3. Place the column in a clean 1.5 or 2.0 ml microcentrifuge
tube. Carefully apply the sample (20–75 µl) directly to the
center of the column. Application of more or less than the
recommended sample volume may decrease column per-
formance.
4. After loading sample, centrifuge the column for 4 minutes
at 1,000 x (g).
5. Following centrifugation, the purified sample is now in
either SSC or Tris buffer. Molecules smaller than the col-
umn’s exclusion limit will be retained by the column (see
Specifications).
6. Properly dispose of the used column.
Micro Bio-Spin ® Chromatography Columns