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| Product Name | Magnetic Beads for PCR Product Purification and Size Selection |
| Catalog No. | MAGBEA-0020 |
| Description | A dual-function carboxyl-modified magnetic bead formulation for PCR product purification and DNA size selection through paramagnetic bead-based size fractionation (SPRI technology). By adjusting the volumetric ratio of bead suspension to sample, specific DNA fragment size ranges are selectively bound, allowing both single-size purification and double-sided size selection for NGS library preparation. The carboxyl surface chemistry utilizes charge-switching and crowding agent effects for tunable, sequence-independent DNA binding based on fragment length. |
| Intended Use | Purification of PCR products, enzymatic reactions, and NGS library fragments; size selection of DNA fragments ranging from 100 bp to 2 kb for next-generation sequencing library construction, targeted sequencing, and long-read sequencing applications; removal of primer-dimers, adapters, and unincorporated nucleotides. |
| Principle / Technology | Carboxyl-functionalized magnetic beads in a PEG/NaCl crowding agent buffer bind DNA in a size-dependent manner based on the volumetric ratio of beads to sample. At low bead-to-sample ratios, only larger DNA fragments are precipitated and captured; at higher ratios, progressively smaller fragments bind. This tunable binding enables left-side selection (removing small fragments), right-side selection (removing large fragments), or double-sided selection (isolating a narrow size range). After magnetic capture and ethanol washing, size-selected DNA is eluted in low-salt buffer or water. |
| Detection Method | Bioanalyzer or TapeStation for size distribution analysis; Qubit fluorometric quantitation; agarose gel electrophoresis; qPCR for adapter-dimer removal verification; NGS library quality metrics (insert size, library yield, sequencing cluster density) |
| Sample Type | PCR amplicons, restriction fragments, NGS library DNA (end-repaired, A-tailed, and adapter-ligated), fragmented genomic DNA, cDNA, and other double-stranded DNA in enzymatic reaction buffers, TE, or nuclease-free water; compatible with standard PCR master mixes and NGS library preparation chemistries |
| Performance Range / Specifications | Bead diameter: 0.5-1 µm; binding capacity: 2-5 µg DNA per 100 µL bead suspension; size selection range: 100 bp-2 kb (tunable by bead:sample ratio); left-side cutoff precision: ±25 bp; right-side cutoff precision: ±50 bp; primer-dimer removal: >99% (<100 bp fragments excluded); DNA recovery: 70-95% (size and ratio dependent); NGS library yield improvement: 2-5× over non-size-selected; magnetic separation: <5 minutes |
| Sensitivity / LOD | PCR product recovery from reactions containing <10 ng input; size selection from DNA amounts as low as 5 ng; effective primer-dimer removal from reactions with >90% undesired short products; single-base resolution of size-cutoff not achievable—expect 25-50 bp cutoff tolerance |
| Specificity | Sequence-independent size-selective binding; no GC-content bias within standard PEG/NaCl system; minimal single-stranded DNA binding under recommended conditions; no enrichment of specific sequence motifs; uniform recovery across diverse DNA sequences; no detectable bead carryover in eluate |
| Reaction Conditions / Protocol | For PCR cleanup (no size selection): add 1.8-2.0 volumes bead suspension to PCR reaction; mix thoroughly; incubate 5-10 minutes at room temperature; place on magnetic rack for 5 minutes or until solution clears; remove supernatant; wash beads twice with 80% ethanol (200 µL each) without disturbing bead pellet; air-dry beads for 5-10 minutes; elute in 10-50 µL TE buffer or water; incubate 2 minutes; separate beads; collect eluate. For size selection: adjust bead volume ratio according to desired cutoff (lower ratio for larger fragments); for double-sided selection, perform sequential binding with two different bead ratios. Total time: 20-40 minutes. |
| Components / Formulation | Carboxyl-modified superparamagnetic beads, suspended at 20-50 mg/mL in a proprietary buffer containing PEG 8000 (10-20%), NaCl (1-2.5 M), Tris-HCl (10 mM, pH 8.0), EDTA (1 mM), and 0.02% sodium azide preservative; supplied as ready-to-use suspension; ethanol for washing not included |
| Storage Conditions | 2-8°C (do not freeze—freezing alters PEG/NaCl phase behavior and bead performance); protect from light; warm to room temperature and vortex thoroughly before use; tightly seal container to prevent evaporation (PEG concentration critical); use within 6 months of opening |
| Shelf Life | 18 months at 2-8°C; PEG/NaCl buffer stable; bead binding properties maintained; size selection cutoff stability verified by periodic calibration with DNA ladder; opened: 6 months with proper handling |
| Package Specifications | 5 mL, 15 mL, 50 mL, 100 mL, 500 mL ready-to-use suspension; sufficient for 50-5000 purifications; bulk OEM packaging; trial sizes for evaluation; supplied in drip-free dispensing bottles |
| Product Form | Opaque white to pale suspension; homogeneous after vigorous vortexing; stable suspension with minimal settling; DO NOT centrifuge—vortex to resuspend settled beads; consistency similar to milk in appearance |
| Quality Control | Per lot: DNA binding capacity, size selection calibration (determination of cutoff ratio for 150 bp and 500 bp fragments using DNA ladder), DNA recovery, primer-dimer removal efficiency, NGS library size selection performance (Bioanalyzer traces), endotoxin, sterility, PEG and NaCl concentration, pH, bead concentration; each lot calibrated with standardized DNA ladder |
| Key Features | Tunable size selection by bead ratio adjustment; single reagent for cleanup and size selection; no columns or gel extraction required; consistent performance across DNA concentrations; validated for all major NGS library prep kits; rapid 20-minute protocol; cost-effective alternative to gel-based size selection; room temperature protocol; compatible with automation |
| Purity | Endotoxin <0.05 EU/mg; DNase free; RNase free; no PCR inhibitors in eluate; PEG and NaCl are molecular biology grade; sodium azide within specification |
| Concentration | 20-50 mg/mL bead suspension in PEG/NaCl buffer; exact concentration per lot; binding capacity 2-5 µg DNA per 100 µL suspension; ready-to-use—do not dilute |
| Activity / Unit Definition | DNA binding: 2-5 µg/100 µL suspension; size selection cutoff precision: ±25 bp (left), ±50 bp (right); primer removal >99%; recovery 70-95%; activity maintained for 18 months |
| Molecular Weight | PEG 8000: ~8,000 Da average molecular weight; beads: submicron carboxyl-functionalized superparamagnetic particles; exact molecular weight not defined for particulate component |
| Source / Origin | Carboxyl-modified superparamagnetic beads are synthetic polymer-magnetite composite; PEG 8000 is synthetic polymer; all buffer components are molecular biology grade, nuclease-free; no animal-derived components; all raw materials traceable |
| pH Range / Optimal pH | Binding buffer pH 7.5-8.5; size selection unaffected by sample pH within 6.5-9.0; elution: nuclease-free water (pH 6-8) or TE (pH 8.0); PEG precipitation efficiency pH-dependent—avoid pH <6 or >9.5; bead integrity: pH 3-12 |
| Shipping Conditions | Ambient temperature acceptable; cold packs for summer; do not freeze; protect from extreme heat (>40°C); standard courier; ambient shipping stable for 2-4 weeks |
| Expiration Date / Stability | 18 months at 2-8°C; real-time stability: binding capacity, size selection cutoff, DNA recovery all within specification at 18 months; accelerated stability (37°C, 14 days) supports shelf life; prevent evaporation—PEG concentration is critical; after opening: 6 months |
| Regulatory / Compliance | For research use only; suitable for NGS library preparation in research and clinical research applications; not for diagnostic procedures; ISO 9001 manufacturing; certificate of analysis per lot |
| Compatibility | Compatible with all major NGS library preparation kits (Illumina, MGI, Thermo Fisher, NEB, Kapa/Roche, Swift); compatible with PCR products in standard buffer systems; compatible with enzymatic reaction buffers (end-repair, A-tailing, ligation); DNA eluted in water, TE, or EB buffer; compatible with 96-well plate format and automated liquid handlers; works with standard magnetic racks and plate magnets |
| Recommended Buffer System | Binding buffer: PEG 8000, NaCl, Tris-HCl pH 8.0, EDTA; ethanol wash: 80% ethanol (freshly prepared); elution: 10 mM Tris-HCl pH 8.0-8.5, 0.1 mM EDTA or nuclease-free water; all components pre-formulated in ready-to-use suspension |
| Application Notes / Precautions | Vortex bead suspension vigorously and warm to room temperature before each use (30 minutes); PEG/NaCl solution is viscous—pipette slowly and accurately; bead-to-sample ratio is critical for reproducible size selection—always use calibrated pipettes; for double-sided selection, remove supernatant completely between binds; over-drying beads causes pellet cracking and reduced recovery; under-drying leaves ethanol that inhibits downstream reactions; elute in pre-warmed buffer for maximum recovery; for samples with high EDTA (>5 mM), supplement PEG/NaCl buffer with additional NaCl; calibrate size selection cutoff for each new lot with DNA ladder |
| Batch-to-Batch Consistency | Size selection cutoff calibrated per lot using standardized DNA ladder; binding capacity CV <12%; primer-dimer removal >99% all lots; DNA recovery CV <10%; PEG and NaCl concentration within ±5% of specification; each lot provided with calibration data for optimal bead ratios |
For research use only, not for clinical use.
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