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| Product Name | Oligo-dT (18) Primer (50 μM) |
| Catalog No. | NATR-HMM-0137 |
| Description | A high-purity, HPLC-purified oligonucleotide consisting of 18 consecutive deoxythymidine residues for the selective priming of polyadenylated messenger RNA in reverse transcription reactions. The primer specifically anneals to the poly(A) tails of eukaryotic mRNA, providing selective cDNA synthesis. |
| Intended Use | Selective first-strand cDNA synthesis from eukaryotic mRNA via reverse transcription for gene expression analysis, cDNA library construction, 3ʹ-RACE (Rapid Amplification of cDNA Ends), and mRNA-seq library preparation. |
| Principle / Technology | The oligo-dT(18) primer hybridizes specifically to the poly(A) tail present at the 3ʹ end of mature eukaryotic mRNAs. Reverse transcriptase extends from the primer in the 5ʹ-to-3ʹ direction along the mRNA template, generating a complementary DNA copy enriched for mRNA sequences and largely excluding ribosomal RNA, transfer RNA, and other non-polyadenylated RNA species. |
| Detection Method | cDNA yield measured by fluorometry or UV spectrophotometry; enrichment of mRNA targets in cDNA verified by RT-qPCR comparing mRNA targets (GAPDH, ACTB) to rRNA (18S, 28S) amplification; cDNA length analysis by Bioanalyzer or alkaline agarose gel electrophoresis |
| Sample Type | Total RNA from eukaryotic cells and tissues; purified eukaryotic mRNA; synthetic RNA with poly(A) tails; samples from organisms with poly(A)+ mRNA |
| Performance Range / Specifications | Primer concentration: 50 μM in nuclease-free water or TE; typical usage: 1–5 μM final concentration (0.25–1.25 μg per 20 μL cDNA synthesis reaction); poly(A) tail annealing temperature: 37–50°C depending on reverse transcriptase thermal optimum; ≥18 consecutive T residues ensure stable annealing |
| Sensitivity / LOD | Reverse transcription from as little as 1 pg of purified mRNA; selective amplification of mRNA targets over rRNA by 100–10,000 fold compared to random hexamer priming; enriches for 3ʹ transcript ends |
| Specificity | Specific annealing to poly(A) tails; no priming from internal poly(A) stretches (>90% of cDNA derived from 3ʹ end of transcripts); >95% HPLC purity (full-length 18-mer); no detectable shorter oligonucleotide contaminants |
| Reaction Conditions / Protocol | Mix RNA, oligo-dT(18) primer (1–2 μL of 50 μM stock), dNTPs, and nuclease-free water; denature at 65°C for 5 minutes; chill on ice; add RT buffer, RNase inhibitor, and reverse transcriptase; incubate at 42–50°C for 30–60 minutes; heat inactivate at 70°C for 10 minutes; proceed to PCR or store at –20°C |
| Components / Formulation | Oligo-dT(18) primer (5ʹ-TTTTTTTTTTTTTTTTTT-3ʹ, sodium salt), HPLC-purified, supplied at 50 μM in nuclease-free water or TE buffer (10 mM Tris-HCl pH 8.0, 1 mM EDTA) |
| Storage Conditions | –20°C for long-term storage; stable at 2–8°C for up to 3 months; avoid repeated freeze-thaw (>30 cycles); aliquot single-use volumes; protect from contamination with RNases |
| Shelf Life | 24 months at –20°C from date of manufacture |
| Package Specifications | 5 μg (1 vial at 50 μM), 25 μg, and 100 μg packaging options |
| Product Form | Ready-to-use liquid primer solution; no heating or reconstitution required |
| Quality Control | Each lot tested: HPLC purity >95% for full-length 18-mer; RT-qPCR with 1 μg HeLa total RNA: GAPDH Ct ≤20; rRNA contamination by RT-qPCR: 18S and 28S Ct >30 (compared to GAPDH Ct <20); no detectable DNase/RNase activity; functional testing with M-MLV and AMV reverse transcriptases |
| Key Features | HPLC-purified for consistent RT priming efficiency; selective mRNA priming reduces rRNA background in cDNA; standard 18-mer length optimizes stability and specificity; minimal lot-to-lot variation for reproducible results; compatible with all major commercial reverse transcriptases |
For research use only, not for clinical use.
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