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Overview
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.
 Bio-Spin® 6 |
 Micro Bio-Spin™ 6 |
Bio-Spin 30 |
Micro Bio-Spin 30 |
 PCR Kleen™ |
|
| 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.
