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| Product Name | In Vitro Transcription Template and Reaction Kit, T7 Polymerase |
| Catalog No. | ATR-CTR-0014 |
| Description | A complete kit for the in vitro synthesis of capped and polyadenylated mRNA from linearized plasmid DNA templates, using T7 RNA polymerase for high-yield transcription suitable for transfection and microinjection applications. |
| Intended Use | Synthesis of functional mRNA for transfection experiments, microinjection into oocytes and embryos, in vitro translation systems, and RNA-based vaccine and therapeutic research. |
| Principle / Technology | T7 bacteriophage RNA polymerase initiates transcription from its cognate promoter sequence with high specificity and processivity; co-transcriptional capping with anti-reverse cap analog (ARCA) ensures proper orientation; poly A tail added by post-transcriptional enzymatic polyadenylation. |
| Detection Method | RNA yield by spectrophotometry and fluorometry (Qubit RNA), RNA integrity by Bioanalyzer or denaturing gel electrophoresis, translation efficiency by in vitro translation assay |
| Sample Type | Linearized plasmid DNA template with T7 promoter |
| Performance Range / Specifications | RNA yield 50–100 µg per 20 µL reaction; >90% full-length transcripts; >80% capped mRNA; poly A tail 150–200 nucleotides |
| Sensitivity / LOD | N/A |
| Specificity | N/A |
| Reaction Conditions / Protocol | Assemble transcription reaction with linearized template DNA, T7 polymerase, NTP mix, cap analog, and buffer; incubate at 37°C for 2 hours; treat with DNase I to remove template; purify mRNA by lithium chloride precipitation or column. |
| Components / Formulation | T7 RNA Polymerase (200 U/µL), 5x Transcription Buffer, NTP mix (ATP, CTP, UTP, GTP), ARCA cap analog, DNase I, nuclease-free water, positive control template |
| Storage Conditions | -20°C; T7 polymerase at -20°C in 50% glycerol (do not freeze-thaw repeatedly) |
| Shelf Life | 12 months at -20°C |
| Package Specifications | Kit for 25 reactions, 100 reactions |
| Product Form | Enzymes, nucleotides, and buffers, RNase free |
| Quality Control | T7 polymerase activity verification, RNase/DNase free, RNA yield from control template, in vitro translation validation |
| Key Features | Complete kit; ARCA cap analog for translation-competent mRNA; DNase treatment included; validated for transfection-grade mRNA synthesis; high yield |
For research use only, not for clinical use.
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