Bio-Rad offers a variety of pre-PCR and post-PCR sample preparation and cleanup columns and prepacked columns; many are also suitable for DNA fragment purification.
Freeze 'N Squeeze DNA Gel Extraction Spin Columns |
Micro Bio-Spin and Bio-Spin Columns | PCR Kleen Spin Columns |
|
Application | Recovery of DNA fragments (50 bp–23 kb) purified from TAE- or TBE-buffered agarose gels | Dye terminator and label cleanup from sequencing reactions; buffer exchange; desalting | Removal of PCR products >200 bp from other reaction mixture components |
Starting Material | ≤700 mg gel slice | 10–100 µl of sequencing reaction mixture | 25–100 µl of PCR or other enzyme reaction |
Format | Spin-column extraction of frozen-thawed gel slices | Prepacked size exclusion spin column | Prepacked size exclusion spin column |
Recovery | 4–23 kb: >80% recovery; 0.5–4 kb: >70% recovery; 50–500 bp: >50% recovery |
>95% of linear DNA >22 bases or bp; 99% retention of unincorporated dye terminators | 50–80% |
Time Required | <10 min | <10 min | <4 min |
Effective, rapid removal of primer-dimers from PCR reactions. Two PCR reaction mixtures were purified with PCR Kleen spin columns. Lane 1, a mixture of 2 µg of a 660 bp DNA fragment with 100 ng of a 25-mer primer-dimer; lane 3, a mixture of 2 µg of a 660 bp DNA fragment with 500 ng of a 25-mer primer; lane 2, primer-dimer purification; lane 4, primer purification; M, DNA markers. Reaction products and purified samples (10 µl each) were analyzed on a 1.5% agarose gel.
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Packed support | Special grade Bio-Gel® P-6 Gel |
Special grade Bio-Gel P-6 Gel |
Special grade Bio-Gel P-30 Gel |
Special grade Bio-Gel P-30 Gel |
Special grade Size exclusion Gel |
Equilibration buffer | 10 mM Tris, pH 7.4 or SSC buffer* | 10 mM Tris, pH 7.4 or SSC buffer* | 10 mM Tris, pH 7.4 or SSC buffer* | 10 mM Tris, pH 7.4 or SSC buffer* | 10 mM Tris, 1 mM EDTA, pH 7.0 |
Applications Desalting of oligonucleotides >20 bases | • | • | – | – | – |
Labeling reactions: removal of unincorporated nucleotides >20 bases or bp from DNA |
– | – | • | • | – |
Removal of primers and primer-dimers from PCR products >200 bp | – | – | – | – | • |
Buffer exchange (restriction fragments, PCR products, enzyme reactions, sequencing templates) |
• | • | – | • | – |
DNA sequencing reaction mixture cleanup** |
– | – | • | • | – |
Riboprobe cleanup*** | – | – | – | • | – |
Desalting of antibody, enzyme, and protein solutions | • | • | – | • | – |
Purification of proteins of moleular weight >6,000 | • | • | – | – | – |
Purification of proteins of molecular weight >40,000 | – | – | • | • | – |
Bed Volume | 1.1 ml | 0.7 ml | 1.1 ml | 0.7 ml | 0.6 ml |
Retention and recovery | 90% recovery of 20 bases or bp, 99% retention of salts | 90% recovery of 20 bases or bp, 99% retention of salts | 95% recovery of 22 bases or bp, 98% retention of ddNTPs | 95% recovery of 22 bases or bp, 98% retention of ddNTPs | 85% recovery of ≥700bp, 95% retention of primers and primer-dimers |
Molecular weight exclusion limit, globular proteins | 6,000 | 6,000 | 40,000 | 40,000 | 8,000,000 |
Sample volume | 50–100 µl | 10–75 µl | 50–100 µl | 10–75 µl | 25–100 µl |
Centrifuge type | Swinging bucket | Microcentrifuge | Swinging bucket | Microcentrifuge | Microcentrifuge |
Autoclavability | Yes | Yes | Yes | Yes | Yes |
* 150 mM NaCl, 17.5 mM sodium citrate, pH 7.0
** In Tris buffer.
*** In RNase-free Tris buffer.
PCR Kleen columns are prepacked spin columns for purifying PCR products and other DNA molecules >200 bp directly from reaction mixtures.
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