{"product_id":"fast-direct-rt-premix-ung-probe-qrt-pcr","title":"Fast Direct RT Premix-UNG (Probe qRT-PCR)","description":"\u003ch2\u003eProduct Overview\u003c\/h2\u003e\n\u003cp\u003eFast Direct RT Premix-UNG (Probe qRT-PCR)(FG-PF-S25) is a pre-dispensed, freeze-dried bead-form reagent designed for direct target-gene real-time fluorescence detection of RNA (TaqMan probe method) without a nucleic acid extraction or purification step. It contains a genetically engineered, screened fast-amplification reverse transcriptase and DNA polymerase, enabling completion of the PCR reaction within 20–40 minutes. This product exhibits strong tolerance to PCR inhibitors, supporting direct RNA amplification from complex, inhibitor-rich sample matrices without a separate nucleic acid extraction or purification step. The reagent uses a mixed enzyme system of an inhibitor-resistant amplification enzyme and UNG enzyme, together with an optimized buffer system containing dUTP, which not only enables good amplification of the target gene in samples containing inhibitors, but also effectively prevents false-positive results caused by residual PCR products and aerosol contamination. This product is stable at room temperature and can be transported and stored at ambient temperature.\u003c\/p\u003e\n\u003ch2\u003eReagent Composition\u003c\/h2\u003e\n\u003cp\u003eFast Direct RT Premix-UNG (Probe qRT-PCR) (020303) (PF-S25) already contains a fast-amplification DNA polymerase, RTase, TS-UNG, PCR Buffer, MgCl2, dNTPs, stabilizers, lyoprotectants, and other components.\u003c\/p\u003e\n\u003ch2\u003eSpecifications\u003c\/h2\u003e\n\u003ctable border=\"1\" cellpadding=\"6\" cellspacing=\"0\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eProduct Name\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eFast Direct RT Premix-UNG (Probe qRT-PCR) (020303)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eCatalog No.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eFG-PF-S25\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eFormat\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003ePre-dispensed, freeze-dried bead-form reagent\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDetection Method\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eTaqMan probe-based real-time RT-PCR\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eExtraction Required\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eNo — direct amplification from raw sample without nucleic acid extraction\/purification\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eSample Matrix Compatibility\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eFormulated for complex, PCR inhibitor-rich sample matrices for research use. See Instructions for Use for tested input volumes and ratios.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eTotal Reaction Time\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e20–40 min\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eContamination Control\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eUNG\/dUTP carryover-prevention system (TS-UNG, temperature-sensitive)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Conditions\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eCool, dry place or 2°C–8°C; stable at room temperature for transport\/storage\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003ch2\u003eInstructions for Use\u003c\/h2\u003e\n\u003cp\u003e\u003cstrong\u003eStep 1:\u003c\/strong\u003e Distribute the lyophilized beads into 8-tube strips.\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eStep 2:\u003c\/strong\u003e Add template, primers, and probe, and top up to 25 µL with water:\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\u003e25 µL system\u003c\/th\u003e\n\u003cth\u003eFinal concentration\u003c\/th\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003eFast Direct RT Premix-UNG (Probe qRT-PCR)\u003c\/td\u003e\n\u003ctd\u003e——\u003c\/td\u003e\n\u003ctd\u003e——\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e25× Primer-Probe Mix\u003c\/td\u003e\n\u003ctd\u003e1 µL\u003c\/td\u003e\n\u003ctd\u003e1×\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\u003e——\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003eddH2O\u003c\/td\u003e\n\u003ctd\u003eTo 25 µL\u003c\/td\u003e\n\u003ctd\u003e——\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003cp\u003ePrimer concentration is typically 0.2 µM for standard PCR (adjustable 0.2–1 µM). Probe concentration is typically optimized within 0.1–0.3 µM.\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eReference sample input volumes for research use (50 µL reaction), based on internal testing:\u003c\/strong\u003e\u003c\/p\u003e\n\u003ctable border=\"1\" cellpadding=\"6\" cellspacing=\"0\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003cth\u003eSample matrix\u003c\/th\u003e\n\u003cth\u003eInput volume\u003c\/th\u003e\n\u003cth\u003eMaximum input ratio\u003c\/th\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003eWhole blood matrix (anticoagulated)\u003c\/td\u003e\n\u003ctd\u003e2.5 µL\u003c\/td\u003e\n\u003ctd\u003e5%\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003eSwab-based matrix\u003c\/td\u003e\n\u003ctd\u003e10 µL\u003c\/td\u003e\n\u003ctd\u003e20%\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003cp\u003e\u003cem\u003eThese are starting-point reference values for research use; optimal input volume should be determined based on your specific sample type, collection method, and required sensitivity.\u003c\/em\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eStep 3:\u003c\/strong\u003e Cap the tubes, mix and centrifuge, then load onto the instrument.\u003c\/p\u003e\n\u003ch2\u003eReaction Conditions\u003c\/h2\u003e\n\u003cp\u003e\u003cstrong\u003eStandard PCR Program:\u003c\/strong\u003e\u003c\/p\u003e\n\u003ctable border=\"1\" cellpadding=\"6\" cellspacing=\"0\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003cth\u003eStep\u003c\/th\u003e\n\u003cth\u003eTemperature\u003c\/th\u003e\n\u003cth\u003eDuration\u003c\/th\u003e\n\u003cth\u003eCycles\u003c\/th\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003eReverse transcription\u003c\/td\u003e\n\u003ctd\u003e50°C\u003c\/td\u003e\n\u003ctd\u003e10–20 min\u003c\/td\u003e\n\u003ctd\u003e1\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003eDenaturation\u003c\/td\u003e\n\u003ctd\u003e95°C\u003c\/td\u003e\n\u003ctd\u003e1–5 min\u003c\/td\u003e\n\u003ctd\u003e1\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003eDenaturation\u003c\/td\u003e\n\u003ctd\u003e95°C\u003c\/td\u003e\n\u003ctd\u003e10–20 s\u003c\/td\u003e\n\u003ctd\u003e40–50\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003eAnnealing\/Extension\u003c\/td\u003e\n\u003ctd\u003e56–64°C\u003c\/td\u003e\n\u003ctd\u003e20–60 s\u003c\/td\u003e\n\u003ctd\u003e(same 40–50)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003cp\u003e\u003cstrong\u003eFast PCR Program:\u003c\/strong\u003e\u003c\/p\u003e\n\u003ctable border=\"1\" cellpadding=\"6\" cellspacing=\"0\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003cth\u003eStep\u003c\/th\u003e\n\u003cth\u003eTemperature\u003c\/th\u003e\n\u003cth\u003eDuration\u003c\/th\u003e\n\u003cth\u003eCycles\u003c\/th\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003eReverse transcription\u003c\/td\u003e\n\u003ctd\u003e50°C\u003c\/td\u003e\n\u003ctd\u003e5 min\u003c\/td\u003e\n\u003ctd\u003e1\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003eDenaturation\u003c\/td\u003e\n\u003ctd\u003e95°C\u003c\/td\u003e\n\u003ctd\u003e30 s\u003c\/td\u003e\n\u003ctd\u003e1\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003eDenaturation\u003c\/td\u003e\n\u003ctd\u003e95°C\u003c\/td\u003e\n\u003ctd\u003e1–3 s\u003c\/td\u003e\n\u003ctd\u003e40–45\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003eAnnealing\/Extension\u003c\/td\u003e\n\u003ctd\u003e56–64°C\u003c\/td\u003e\n\u003ctd\u003e3–20 s\u003c\/td\u003e\n\u003ctd\u003e(same 40–45)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003cp\u003e\u003cem\u003eThe temperature-sensitive TS-UNG (DG) enzyme is active at room temperature and is inactivated during the reverse transcription step.\u003c\/em\u003e\u003c\/p\u003e\n\u003ch2\u003eTechnical Notes\u003c\/h2\u003e\n\u003cul\u003e\n\u003cli\u003eThe fast DNA polymerase amplification rate is no less than 1 kb\/10 s. Reaction conditions should be optimized for the specific fast PCR instrument used, since heating\/cooling rates and thermal conductivity vary by instrument.\u003c\/li\u003e\n\u003cli\u003eIf fluorescence values are too low or amplification inhibition is evident, reduce the sample input volume or dilute the sample before use.\u003c\/li\u003e\n\u003cli\u003eSample collection should follow clinical standard operating procedures; freshly collected samples help avoid nucleic acid degradation.\u003c\/li\u003e\n\u003cli\u003eFor primers with lower annealing temperatures or amplicons longer than 200 bp, a three-step protocol is recommended.\u003c\/li\u003e\n\u003cli\u003eDifferent target genes vary in their utilization efficiency of dUTP and sensitivity to UNG enzyme; if using the UNG system reduces detection sensitivity, contact our company for technical support to adjust the reaction system.\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\u003eDo I need to extract RNA before using this reagent?\u003c\/span\u003e \u003cspan class=\"fg-faq-icon\"\u003e+\u003c\/span\u003e \u003c\/button\u003e\n\u003cdiv class=\"fg-faq-answer\"\u003e\n\u003cp\u003eNo. This reagent is designed for direct target-gene amplification from raw, complex sample matrices without a separate nucleic acid extraction or purification step, thanks to its inhibitor-tolerant enzyme system. See the Instructions for Use section for reference sample matrices and tested input volumes.\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 detection method does this product use?\u003c\/span\u003e \u003cspan class=\"fg-faq-icon\"\u003e+\u003c\/span\u003e \u003c\/button\u003e\n\u003cdiv class=\"fg-faq-answer\"\u003e\n\u003cp\u003eIt uses the TaqMan probe method for real-time fluorescence detection of RNA via RT-PCR.\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 long does the full reaction take?\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 complete PCR reaction can be finished within 20-40 minutes, depending on whether the Standard or Fast PCR program is used.\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 sample matrices have been tested, and how much sample should I add?\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 a 50 µL reaction, internal reference testing used 2.5 µL of a whole blood matrix (up to 5% of reaction volume) or 10 µL of a swab-based matrix (up to 20% of reaction volume). These are starting points for research use — optimal input should be validated based on your specific sample type, collection method, and required sensitivity.\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 does this product prevent carryover contamination?\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 reagent uses a UNG enzyme and dUTP-containing buffer system. The temperature-sensitive TS-UNG enzyme is active at room temperature to degrade contaminating amplicons, then is inactivated during the reverse transcription step.\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 this product be stored and transported?\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 the freeze-dried reagent in a cool, dry place or at 2°C–8°C. The product is stable at room temperature and can be transported and stored at ambient temperature.\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 primer and probe concentrations should I use?\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 standard PCR programs, a final primer concentration of 0.2 µM typically works well (adjustable within 0.2-1 µM if needed), with probe concentration optimized within 0.1-0.3 µM. For fast PCR programs, increasing primer\/probe concentration may improve results.\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's the difference between the Standard and Fast PCR programs?\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 Standard program uses a 10-20 min reverse transcription step and 40-50 amplification cycles at 10-20 s per denaturation step. The Fast program shortens reverse transcription to 5 min and denaturation to 1-3 s per cycle, completing 40-45 cycles more quickly. Instrument-specific optimization is recommended for fast protocols.\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 if using the UNG system reduces my detection sensitivity?\u003c\/span\u003e \u003cspan class=\"fg-faq-icon\"\u003e+\u003c\/span\u003e \u003c\/button\u003e\n\u003cdiv class=\"fg-faq-answer\"\u003e\n\u003cp\u003eDifferent target genes vary in dUTP utilization efficiency and UNG sensitivity. If sensitivity is reduced, the reaction system should be adjusted and optimized — contact our technical support team for guidance.\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 a three-step PCR protocol ever needed?\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. For primers with lower annealing temperatures, or when amplifying fragments longer than 200 bp, a three-step protocol is recommended instead of the standard two-step cycling.\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":48635380859092,"sku":"FG-PF-S25-100","price":459.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0634\/0912\/7636\/files\/FastDirectRTPremix-UNG_ProbeqRT-PCR.png?v=1788852511","url":"https:\/\/firegene.com\/products\/fast-direct-rt-premix-ung-probe-qrt-pcr","provider":"FireGene","version":"1.0","type":"link"}