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| Product Name | Multiplex Ligation-Dependent Probe Amplification (MLPA) Probe Synthesis Service |
| Catalog No. | PRIPRO-0030 |
| Description | Custom synthesis of MLPA probe pairs for multiplex detection of copy number variations (CNVs), gene deletions/duplications, and methylation status analysis. Each target requires two hemi-probes that hybridize to adjacent sequences and are ligated by a thermostable ligase only when both probes are perfectly matched. Ligated probes are amplified by PCR using universal primers with one primer fluorescently labeled, separated by capillary electrophoresis for relative quantification. |
| Intended Use | Copy number variation (CNV) detection; gene deletion and duplication analysis; DNA methylation analysis (MS-MLPA); aneuploidy screening; diagnostic genetic testing for microdeletion/microduplication syndromes; mRNA expression profiling. |
| Principle / Technology | Two hemi-probes per target: left probe oligo (LPO) contains universal forward primer sequence + target-specific sequence; right probe oligo (RPO) contains target-specific sequence + universal reverse primer sequence + stuffer sequence for length discrimination; probes ligate only when both hybridize adjacently; ligated probes PCR-amplified with fluorescent universal primers; CE separation resolves targets by stuffer length (130-480 nt) |
| Detection Method | Customer provides target sequences for each genomic locus of interest (100-200 bp); probes designed with unique stuffer lengths for multiplex resolution; supplied as probe mix with all LPOs and RPOs; standard scale 25 nmol each, PAGE-purified |
| Sample Type | Customer provides genomic coordinates (GRCh38/hg38 or GRCh37/hg19) and specific target sequence for each locus; indicate if MS-MLPA (methylation-specific) probes required |
| Components / Formulation | Custom probe mix: left probe oligos + right probe oligos for each target; universal PCR primers (optional, FAM or HEX labeled); probe mix supplied as 0.2 µM each oligo concentration in TE buffer |
| Storage Conditions | Lyophilized or in TE buffer at -20 °C; protect fluorescent primers from light |
| Shelf Life | Lyophilized: 24 months at -20 °C; in TE solution: 12 months at -20 °C |
| Package Specifications | Custom service; probe mix for 20-50 targets; synthesis scale 25-100 nmol per oligo; 5′-phosphorylation of RPO included |
| Product Form | Lyophilized oligonucleotides or premixed probe solution in TE buffer |
| Key Features | Custom probe design for each locus; PAGE-purified for CE compatibility; 5′-phosphorylation of RPOs included; unique stuffer sequences for up to 50-plex detection; universal primer sequences compatible with commercial MLPA kits; quality control by capillary electrophoresis of ligated probes |
| Purity | PAGE-purified: >95% full-length; each probe verified by ESI-MS; RPO phosphorylation >95% efficiency confirmed spectrophotometrically |
| Concentration | As specified in synthesis report; typically lyophilized or at 100 µM in TE |
| Activity / Unit Definition | Primer efficiency 90–110% in qPCR; Tm within ±2 °C of calculated value |
| Molecular Weight | As specified per MALDI-TOF certificate; within 0.1% of theoretical mass |
| Source / Origin | Solid-phase phosphoramidite chemical synthesis |
| pH Range / Optimal pH | Stable in TE buffer pH 7.0–8.0; avoid acidic conditions causing depurination |
| Shipping Conditions | Ambient temperature; lyophilized probes stable at RT 2 weeks; fluorescent primers ship on cold pack protected from light |
| Expiration Date / Stability | 24 months lyophilized at -20 °C; 12 months in solution at -20 °C; protect fluorescent materials from light |
| Regulatory / Compliance | For laboratory and research use only; RUO; for diagnostic use, additional validation required per local regulations; ISO 9001 |
| Compatibility | Compatible with standard PCR, qPCR, and sequencing platforms |
| Recommended Buffer System | Resuspend in nuclease-free water or TE buffer pH 8.0 |
| Application Notes / Precautions | Protect fluorescent probes from light; avoid repeated freeze-thaw cycles; aliquot for single use |
| Batch-to-Batch Consistency | MALDI-TOF mass confirmation per lot; qPCR performance within 0.5 Ct of reference lot |
For research use only, not for clinical use.
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