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| Product Name | PCR Cleanup and Purification Kit |
| Catalog No. | NATR-HMM-0120 |
| Description | A silica membrane spin column-based kit for the rapid purification of DNA from PCR reactions and other enzymatic reaction mixtures. The kit removes primers, nucleotides, enzymes, mineral oil, salts, and other reaction components while recovering double-stranded DNA fragments for downstream use. |
| Intended Use | Purification of PCR products prior to cloning, sequencing, labeling, restriction digestion, microinjection, and microarray spotting; removal of unincorporated primers, primer-dimers, and dNTPs from amplification reactions. |
| Principle / Technology | DNA binds to the silica membrane in the presence of high concentrations of chaotropic salts at an optimized pH. Contaminants including primers, enzymes, dNTPs, and salts are washed away with an ethanol-containing buffer. Pure DNA is eluted under low-ionic-strength conditions in a small volume of buffer or nuclease-free water. |
| Detection Method | UV spectrophotometry for DNA concentration and purity; agarose gel electrophoresis to confirm size and absence of primer-dimers; ABI sequencing for purification quality verification |
| Sample Type | PCR amplification products; restriction digestion products; kinase, phosphatase, fill-in, and other enzymatic reaction mixtures; DNA solutions requiring buffer exchange or concentration |
| Performance Range / Specifications | DNA fragment binding range: 100 bp to 10 kb (standard protocol); <100 bp fragments may be recovered with adjusted buffer conditions; recovery: 80–95% for fragments >200 bp; binding capacity: 10 μg DNA per column; elution volume: 10–50 μL |
| Sensitivity / LOD | Effectively removes >98% of primers (<40-mer) and >99% of unincorporated dNTPs; DNA recovered from as little as 50 ng input |
| Specificity | Selective removal of primers <40 nt and primer-dimers; double-stranded DNA recovery >80%; no detectable carryover of polymerase, restriction enzymes, or reaction buffers |
| Reaction Conditions / Protocol | Add 5 volumes of binding buffer to 1 volume of PCR reaction (or enzymatic reaction), mix thoroughly, transfer to spin column, centrifuge 30–60 seconds, wash with 750 μL wash buffer, centrifuge, dry spin 1–2 minutes, elute with 10–50 μL elution buffer or nuclease-free water (pH 7.0–8.5), incubate 1 minute, centrifuge 1 minute; total protocol time approximately 5–10 minutes |
| Components / Formulation | Binding buffer (guanidine hydrochloride, Tris-HCl, isopropanol), pH indicator dye in binding buffer, wash buffer concentrate (Tris-HCl, NaCl, EDTA, ethanol), elution buffer (10 mM Tris-HCl pH 8.5), silica membrane spin columns with collection tubes, loading dye for agarose gel loading (optional) |
| Storage Conditions | Room temperature (15–25°C); spin columns should be kept sealed in pouch until use; tightly seal binding buffer after use |
| Shelf Life | 24 months from date of manufacture at room temperature |
| Package Specifications | 50, 100, and 250 preparation kits; all columns and buffers included |
| Product Form | Liquid buffers; silica membrane spin columns; pH indicator ensures correct binding conditions |
| Quality Control | Each lot tested: >80% recovery of 200 bp PCR fragment; removal of unincorporated primers and dNTPs >98% verified by gel; purified DNA sequenced with Phred Q20 >700 bases; no co-purification of PCR inhibitors; endotoxin <0.01 EU/μg |
| Key Features | Color-indicator in binding buffer confirms pH for optimal DNA binding; rapid 5-minute protocol; direct scaling to 96-well vacuum format for high throughput; purified DNA ready for sequencing and cloning; concentrated elution option for low-yield PCRs |
For research use only, not for clinical use.
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