{"product_id":"neoscript-rtase-1st-strand-cdna-synthesis-kit","title":"Neoscript RTase 1st Strand cDNA Synthesis Kit","description":"\u003ch2\u003eProduct Overview\u003c\/h2\u003e\n\u003cp\u003eThis kit provides all reagents required for first-strand cDNA synthesis. Neoscript RTase is a reverse transcriptase derived from the Moloney murine leukemia virus (M-MLV) gene, obtained through mutation screening and recombinant expression in E. coli. The RNase H activity of this enzyme has been removed, giving it higher temperature tolerance and making it suitable for high-temperature reverse transcription. This helps eliminate the adverse effects of RNA secondary structure and non-specific factors on cDNA synthesis, and provides greater stability and reverse transcription capability. It can synthesize first-strand cDNA fragments up to 12 kb in length.\u003c\/p\u003e\n\u003ch2\u003eKit Components\u003c\/h2\u003e\n\u003ctable border=\"1\" cellpadding=\"6\" cellspacing=\"0\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003cth\u003eComponent\u003c\/th\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e200 U\/µL Neoscript RTase\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e5× First-Strand Buffer\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e40 U\/µL RNase Inhibitor\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003eOligo(dT)18 Primer (50 µM)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003eRandom 6 mers (50 µM)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e10 mM dNTP Mix\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003ch2\u003eSpecifications\u003c\/h2\u003e\n\u003ctable border=\"1\" cellpadding=\"6\" cellspacing=\"0\" style=\"width: 100.035%;\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 24.8502%;\"\u003e\u003cstrong\u003eProduct Name\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 75.2555%;\"\u003eNeoscript RTase 1st Strand cDNA Synthesis Kit\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 24.8502%;\"\u003e\u003cstrong\u003eCatalog No.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 75.2555%;\"\u003eFG-PM05\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 24.8502%;\"\u003e\u003cstrong\u003eEnzyme Source\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 75.2555%;\"\u003eM-MLV reverse transcriptase, RNase H activity removed, recombinantly expressed in E. coli\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 24.8502%;\"\u003e\u003cstrong\u003eMaximum cDNA Length\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 75.2555%;\"\u003eUp to 12 kb\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 24.8502%;\"\u003e\u003cstrong\u003eOptimal RT Temperature Range\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 75.2555%;\"\u003e42°C–55°C\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 24.8502%;\"\u003e\u003cstrong\u003eUnit Definition\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 75.2555%;\"\u003e1 U = amount of enzyme required to incorporate 1 nmol of dTTP within 10 min at 37°C, using poly(A)·Oligo(dT)25 as template\/primer\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 24.8502%;\"\u003e\u003cstrong\u003ePurity\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 75.2555%;\"\u003eSDS-PAGE purity \u0026gt; 98%\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 24.8502%;\"\u003e\u003cstrong\u003eKey Applications\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 75.2555%;\"\u003eFirst-strand cDNA synthesis, PCR\/qPCR template preparation, cDNA library construction, 3′ and 5′ RACE\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 24.8502%;\"\u003e\u003cstrong\u003eStorage Conditions\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 75.2555%;\"\u003e-20 ± 5°C; mix well before use; avoid repeated freeze-thaw cycles\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003ch2\u003eProtocol Overview\u003c\/h2\u003e\n\u003cp\u003e\u003cstrong\u003eStep 1 — Primer annealing setup (10 µL):\u003c\/strong\u003e\u003c\/p\u003e\n\u003ctable border=\"1\" cellpadding=\"6\" cellspacing=\"0\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003cth\u003eReagent\u003c\/th\u003e\n\u003cth\u003e20 µL system\u003c\/th\u003e\n\u003cth\u003eFinal concentration\u003c\/th\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003eOligo(dT)18 Primer (50 µM)\u003c\/td\u003e\n\u003ctd\u003e1 µL\u003c\/td\u003e\n\u003ctd\u003e2.5 µM\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003eOr Random 6 mers (50 µM)\u003c\/td\u003e\n\u003ctd\u003e1 µL (0.4–2 µL)\u003c\/td\u003e\n\u003ctd\u003e2.5 µM (1–5 µM)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003eOr Gene-specific Primer (10 µM)\u003c\/td\u003e\n\u003ctd\u003e0.2–2 µL\u003c\/td\u003e\n\u003ctd\u003e0.1–1 µM\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e10 mM dNTP\u003c\/td\u003e\n\u003ctd\u003e1 µL\u003c\/td\u003e\n\u003ctd\u003e500 µM\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003eTemplate RNA\u003c\/td\u003e\n\u003ctd\u003e——\u003c\/td\u003e\n\u003ctd\u003eTotal RNA ≤ 5 µg; mRNA ≤ 1 µg\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003eRNase-free dH2O\u003c\/td\u003e\n\u003ctd\u003eTo 10 µL\u003c\/td\u003e\n\u003ctd\u003e——\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003cp\u003eHeat at 65°C for 5 min, then cool rapidly on ice for 2 min.\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eStep 2 — Reverse transcription setup (add to reach 20 µL total):\u003c\/strong\u003e\u003c\/p\u003e\n\u003ctable border=\"1\" cellpadding=\"6\" cellspacing=\"0\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003cth\u003eReagent\u003c\/th\u003e\n\u003cth\u003e20 µL system\u003c\/th\u003e\n\u003cth\u003eFinal concentration\u003c\/th\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e5× First-Strand Buffer\u003c\/td\u003e\n\u003ctd\u003e4 µL\u003c\/td\u003e\n\u003ctd\u003e1×\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e200 U\/µL Neoscript RTase\u003c\/td\u003e\n\u003ctd\u003e1 µL\u003c\/td\u003e\n\u003ctd\u003e10 U\/µL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e40 U\/µL RNase Inhibitor\u003c\/td\u003e\n\u003ctd\u003e0.5 µL\u003c\/td\u003e\n\u003ctd\u003e1 U\/µL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003eRNase-free dH2O\u003c\/td\u003e\n\u003ctd\u003eTo 20 µL\u003c\/td\u003e\n\u003ctd\u003e——\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003cp\u003e\u003cstrong\u003eStep 3 — Incubation:\u003c\/strong\u003e\u003c\/p\u003e\n\u003cul\u003e\n\u003cli\u003eIf using Random Primers: 25°C for 10 min, then 50°C for 30–60 min\u003c\/li\u003e\n\u003cli\u003eIf using Oligo dT or a gene-specific primer: 50°C for 30–60 min\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003e\u003cstrong\u003eStep 4 — Reaction termination:\u003c\/strong\u003e Heat at 95°C for 5 min (or 70°C for 15 min for long-fragment applications) to inactivate Neoscript RTase.\u003c\/p\u003e\n\u003cp\u003eThe reverse transcription product can be used directly for PCR and real-time qPCR, or stored long-term at -20°C.\u003c\/p\u003e\n\u003ch2\u003eTechnical Notes\u003c\/h2\u003e\n\u003cul\u003e\n\u003cli\u003eHigh-purity, intact RNA template is critical for high-quality, full-length cDNA synthesis. Trace genomic DNA contamination may be co-amplified with the target gene; DNase I pretreatment is recommended if genomic DNA-free amplification is required.\u003c\/li\u003e\n\u003cli\u003eAdding RNase Inhibitor effectively suppresses nuclease-mediated degradation of the RNA template.\u003c\/li\u003e\n\u003cli\u003eFor long-fragment amplification, inactivating the reverse transcriptase at 70°C for 15 min (instead of 95°C) helps prevent damage to the cDNA structure.\u003c\/li\u003e\n\u003cli\u003eIf amplified band intensity is weak, consider increasing RNA template and primer concentrations.\u003c\/li\u003e\n\u003cli\u003eNeoscript RTase is suitable for reverse transcription in the 42–55°C range; increasing the RT temperature can improve specificity when amplifying RNA with complex secondary structure.\u003c\/li\u003e\n\u003cli\u003eSuitable for cDNA library construction and 3′\/5′ RACE.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cdiv class=\"fg-faq-section\" style=\"max-width: 900px; margin: 48px auto; padding: 0 16px;\"\u003e\n\u003ch2 style=\"font-size: 28px; margin-bottom: 24px;\"\u003eFrequently Asked Questions\u003c\/h2\u003e\n\u003cdiv class=\"fg-faq-item\" style=\"border-bottom: 1px solid #e0e0e0;\"\u003e\n\u003cbutton class=\"fg-faq-question\"\u003e \u003cspan\u003eWhat is Neoscript RTase derived from?\u003c\/span\u003e \u003cspan class=\"fg-faq-icon\"\u003e+\u003c\/span\u003e \u003c\/button\u003e\n\u003cdiv class=\"fg-faq-answer\"\u003e\n\u003cp\u003eNeoscript RTase is a reverse transcriptase derived from the Moloney murine leukemia virus (M-MLV) gene, obtained through mutation screening and recombinant expression in E. coli, with its RNase H activity removed.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"fg-faq-item\" style=\"border-bottom: 1px solid #e0e0e0;\"\u003e\n\u003cbutton class=\"fg-faq-question\"\u003e \u003cspan\u003eWhat is the maximum cDNA fragment length this kit can synthesize?\u003c\/span\u003e \u003cspan class=\"fg-faq-icon\"\u003e+\u003c\/span\u003e \u003c\/button\u003e\n\u003cdiv class=\"fg-faq-answer\"\u003e\n\u003cp\u003eThe kit can synthesize first-strand cDNA fragments up to 12 kb in length.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"fg-faq-item\" style=\"border-bottom: 1px solid #e0e0e0;\"\u003e\n\u003cbutton class=\"fg-faq-question\"\u003e \u003cspan\u003eWhat temperature range is recommended for reverse transcription?\u003c\/span\u003e \u003cspan class=\"fg-faq-icon\"\u003e+\u003c\/span\u003e \u003c\/button\u003e\n\u003cdiv class=\"fg-faq-answer\"\u003e\n\u003cp\u003eNeoscript RTase performs well across 42°C–55°C. Increasing the reaction temperature within this range can improve specificity when reverse transcribing RNA templates with complex secondary structure.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"fg-faq-item\" style=\"border-bottom: 1px solid #e0e0e0;\"\u003e\n\u003cbutton class=\"fg-faq-question\"\u003e \u003cspan\u003eCan genomic DNA contamination affect my results?\u003c\/span\u003e \u003cspan class=\"fg-faq-icon\"\u003e+\u003c\/span\u003e \u003c\/button\u003e\n\u003cdiv class=\"fg-faq-answer\"\u003e\n\u003cp\u003eTrace genomic DNA introduced during RNA template preparation may be co-amplified with the target gene. If your downstream application requires complete absence of genomic DNA, DNase I pretreatment of the RNA template is recommended.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"fg-faq-item\" style=\"border-bottom: 1px solid #e0e0e0;\"\u003e\n\u003cbutton class=\"fg-faq-question\"\u003e \u003cspan\u003eShould I use Oligo(dT), random hexamers, or a gene-specific primer?\u003c\/span\u003e \u003cspan class=\"fg-faq-icon\"\u003e+\u003c\/span\u003e \u003c\/button\u003e\n\u003cdiv class=\"fg-faq-answer\"\u003e\n\u003cp\u003eFor cDNA synthesis under 2 kb, use 1–2 µL of Random 6 mers; for synthesis over 2 kb, use 0.4–1 µL. A gene-specific primer can also be used at a final concentration of 0.1–1 µM, depending on your target and downstream application.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"fg-faq-item\" style=\"border-bottom: 1px solid #e0e0e0;\"\u003e\n\u003cbutton class=\"fg-faq-question\"\u003e \u003cspan\u003eHow is enzyme activity (unit) defined for this product?\u003c\/span\u003e \u003cspan class=\"fg-faq-icon\"\u003e+\u003c\/span\u003e \u003c\/button\u003e\n\u003cdiv class=\"fg-faq-answer\"\u003e\n\u003cp\u003eOne unit (U) is defined as the amount of enzyme required to incorporate 1 nmol of dTTP within 10 minutes at 37°C, using poly(A)·Oligo(dT)25 as the template\/primer.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"fg-faq-item\" style=\"border-bottom: 1px solid #e0e0e0;\"\u003e\n\u003cbutton class=\"fg-faq-question\"\u003e \u003cspan\u003eWhat purity standard does the enzyme meet?\u003c\/span\u003e \u003cspan class=\"fg-faq-icon\"\u003e+\u003c\/span\u003e \u003c\/button\u003e\n\u003cdiv class=\"fg-faq-answer\"\u003e\n\u003cp\u003eNeoscript RTase meets SDS-PAGE purity greater than 98%, with quality control testing for amplification sensitivity, lot-to-lot variation, stability, and absence of exogenous nuclease contamination.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"fg-faq-item\" style=\"border-bottom: 1px solid #e0e0e0;\"\u003e\n\u003cbutton class=\"fg-faq-question\"\u003e \u003cspan\u003eHow should the kit be stored?\u003c\/span\u003e \u003cspan class=\"fg-faq-icon\"\u003e+\u003c\/span\u003e \u003c\/button\u003e\n\u003cdiv class=\"fg-faq-answer\"\u003e\n\u003cp\u003eStore at -20 ± 5°C. Mix well before use and avoid repeated freeze-thaw cycles.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"fg-faq-item\" style=\"border-bottom: 1px solid #e0e0e0;\"\u003e\n\u003cbutton class=\"fg-faq-question\"\u003e \u003cspan\u003eIs this kit suitable for RACE or cDNA library construction?\u003c\/span\u003e \u003cspan class=\"fg-faq-icon\"\u003e+\u003c\/span\u003e \u003c\/button\u003e\n\u003cdiv class=\"fg-faq-answer\"\u003e\n\u003cp\u003eYes, Neoscript RTase is suitable for cDNA library construction as well as 3′ and 5′ RACE (Rapid Amplification of cDNA Ends).\u003c\/p\u003e\n\u003c\/div\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"fg-faq-item\" style=\"border-bottom: 1px solid #e0e0e0;\"\u003e\n\u003cbutton class=\"fg-faq-question\"\u003e \u003cspan\u003eHow do I inactivate the reverse transcriptase after the reaction?\u003c\/span\u003e \u003cspan class=\"fg-faq-icon\"\u003e+\u003c\/span\u003e \u003c\/button\u003e\n\u003cdiv class=\"fg-faq-answer\"\u003e\n\u003cp\u003eHeat at 95°C for 5 min to terminate the reaction. For long-fragment applications, heating at 70°C for 15 min instead is recommended to avoid damaging the cDNA structure.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003c\/div\u003e\n\u003c\/div\u003e\n\u003cstyle\u003e\n  .fg-faq-question {\n    width: 100%;\n    display: flex;\n    justify-content: space-between;\n    align-items: center;\n    background: none;\n    border: none;\n    text-align: left;\n    padding: 18px 4px;\n    font-size: 16px;\n    font-weight: 600;\n    cursor: pointer;\n    color: #1a1a1a;\n  }\n  .fg-faq-icon {\n    font-size: 20px;\n    font-weight: 400;\n    margin-left: 12px;\n    flex-shrink: 0;\n    transition: transform 0.2s ease;\n  }\n  .fg-faq-question[aria-expanded=\"true\"] .fg-faq-icon {\n    transform: rotate(45deg);\n  }\n  .fg-faq-answer {\n    max-height: 0;\n    overflow: hidden;\n    transition: max-height 0.25s ease;\n    padding: 0 4px;\n  }\n  .fg-faq-answer p {\n    padding-bottom: 18px;\n    margin: 0;\n    color: #4a4a4a;\n    line-height: 1.6;\n  }\n\u003c\/style\u003e\n\u003cp\u003e \u003c\/p\u003e","brand":"FireGene","offers":[{"title":"100 rxns","offer_id":48635367227604,"sku":"FG-PM05","price":479.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0634\/0912\/7636\/files\/NeoscriptRTase1stStrandcDNASynthesisKit.png?v=1788852036","url":"https:\/\/firegene.com\/products\/neoscript-rtase-1st-strand-cdna-synthesis-kit","provider":"FireGene","version":"1.0","type":"link"}