{"product_id":"rt-qpcr-easytm-i-one-step-sybr-green-i","title":"RT-qPCR EasyTM  I (One Step)-SYBR Green I","description":"\u003ch2\u003eDescription\u003c\/h2\u003e\n\u003cp\u003eRT-qPCR EasyTM (One Step)-SYBR Green Iuses a unique system to effectively combine RT and Real Time PCR reactions, which can quickly complete the quantitative detection of genes. The product has strong tolerance, high specificity and stability. The product contains the company's unique Foregene HotStar Taq DNA Polymerase, which has the advantages of faster amplification (2Kb\/min), high amplification efficiency, strong specific amplification ability, and low mismatch rate compared with ordinary Taq enzymes. It is used here for Real Time RT-PCR to reduce non-specific amplification and improve the accuracy of PCR.\u003c\/p\u003e\n\u003ch2\u003eFeatures\u003c\/h2\u003e\n\u003cul\u003e\n\u003cli\u003e\n\u003cp\u003eOne-step kit makes reverse transcription and qPCR two reactions in the same tube, only need to add template RNA, specific PCR primers and RNase-Free ddH2O.\u003c\/p\u003e\n\u003c\/li\u003e\n\u003cli\u003e\n\u003cp\u003eThe kit can quickly and efficiently quantitatively analyze viral RNA or trace RNA.\u003c\/p\u003e\n\u003c\/li\u003e\n\u003cli\u003e\n\u003cp\u003eThe kit uses a unique Foregene reverse transcription reagent and Foregene HotStar Taq DNA Polymerase combined with a unique reaction system to effectively improve the amplification efficiency and specificity of the reaction.\u003c\/p\u003e\n\u003c\/li\u003e\n\u003cli\u003e\n\u003cp\u003eThe optimized reaction system makes the reaction have higher detection sensitivity, stronger thermal stability, and better tolerance.\u003c\/p\u003e\n\u003c\/li\u003e\n\u003cli\u003e\n\u003cp\u003eRT-qPCR EasyTM (One Step)-SYBR Green I kit comes with ROX internal reference dye, which can be used to eliminate signal background and signal errors between wells, which is convenient for customers to use in different models of quantitative PCR instruments.\u003c\/p\u003e\n\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003ch2\u003eKit Contents\u003cbr\u003e\n\u003c\/h2\u003e\n\u003ctable border=\"1\" style=\"width: 90%; border-collapse: collapse;\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"2\" style=\"width: 33.3333%; height: 3px; text-align: center; vertical-align: middle;\"\u003eKit components\u003c\/td\u003e\n\u003ctd style=\"width: 33.3333%; height: 3px; text-align: center; vertical-align: middle;\"\u003eRT-02111\u003c\/td\u003e\n\u003ctd style=\"width: 33.3333%; height: 3px; text-align: center; vertical-align: middle;\"\u003eRT-02112\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 33.3333%; height: 3px; text-align: center; vertical-align: middle;\"\u003e100T (20μl system)\u003c\/td\u003e\n\u003ctd style=\"width: 33.3333%; height: 3px; text-align: center; vertical-align: middle;\"\u003e500T (20μl system)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 33.3333%; height: 3px; text-align: center; vertical-align: middle;\"\u003e2× RT-qPCR EasyTM Mix-SYBR\u003c\/td\u003e\n\u003ctd style=\"width: 33.3333%; height: 3px; text-align: center; vertical-align: middle;\"\u003e1ml\u003c\/td\u003e\n\u003ctd style=\"width: 33.3333%; height: 3px; text-align: center; vertical-align: middle;\"\u003e1.7ml×3\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 33.3333%; height: 3px; text-align: center; vertical-align: middle;\"\u003e50× ROX Reference Dye\u003c\/td\u003e\n\u003ctd style=\"width: 33.3333%; height: 3px; text-align: center; vertical-align: middle;\"\u003e200μl\u003c\/td\u003e\n\u003ctd style=\"width: 33.3333%; height: 3px; text-align: center; vertical-align: middle;\"\u003e1ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 33.3333%; height: 3px; text-align: center; vertical-align: middle;\"\u003eRNase-Free ddH2O\u003c\/td\u003e\n\u003ctd style=\"width: 33.3333%; height: 3px; text-align: center; vertical-align: middle;\"\u003e1.7ml\u003c\/td\u003e\n\u003ctd style=\"width: 33.3333%; height: 3px; text-align: center; vertical-align: middle;\"\u003e1.7ml×3\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 33.3333%; text-align: center; vertical-align: middle;\"\u003eInstruction Manual\u003c\/td\u003e\n\u003ctd style=\"width: 33.3333%; text-align: center; vertical-align: middle;\"\u003e1 piece\u003c\/td\u003e\n\u003ctd style=\"width: 33.3333%; text-align: center; vertical-align: middle;\"\u003e1 piece\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003ch2\u003eKit component information\u003c\/h2\u003e\n\u003cp\u003e2× RT-qPCR EasyTM Mix-SYBR:Foregene Reverse Transcriptase, RNase Inhibitor, Foregene HotStar Taq DNA Polymerase, optimized ratio of dNTPs, SYBR GreenI, Mg2+, stabilizers, enhancers, and optimizers.\u003c\/p\u003e\n\u003cp\u003eROX Reference Dye:it is generally used on Real Time PCR amplifiers of companies such as ABI and Stratagene to adjust the difference between PCR tubes and tubes caused by PCR sample loading errors. The concentration of ROX Reference Dye required by different instruments is different, and the user can add it according to the recommended concentration of the instrument.\u003c\/p\u003e\n\u003cp\u003ePrecautions: (Please read the precautions carefully before using the kit)\u003c\/p\u003e\n\u003cp\u003eReagents should avoid repeated freezing and thawing, otherwise the performance of the reagents will decrease or become invalid.\u003c\/p\u003e\n\u003cp\u003eThe reagents in the kit should be protected from light and stored at -20°C.\u003c\/p\u003e\n\u003cp\u003eIt is recommended to use fresh sample extraction or template RNA stored at -80°C (RNA should avoid repeated freezing and thawing).\u003c\/p\u003e\n\u003cp\u003eIn order to avoid RNase contamination, please perform the experiment in the RNase-Free space; the pipette tips and PCR tubes used must be RNase-Free; and wear disposable gloves and masks.\u003c\/p\u003e\n\u003cp\u003eThis kit must be used with specific primers for experiments. Please select the specific primers for the gene to be amplified according to the needs of the experiment.\u003c\/p\u003e\n\u003cp\u003eBefore use, put 2×RT-qPCR EasyTM Mix-SYBR I on ice to completely melt, flick and mix well before use; the preparation of the system should be operated on an ice bath to improve the performance of the kit and increase PCR amplification The specificity.\u003c\/p\u003e\n\u003cp\u003e2×RT-qPCR EasyTM Mix-SYBR contains SYBR Green I, please avoid strong light.\u003c\/p\u003e\n\u003ch2\u003eTransport and storage conditions\u003cbr\u003e\n\u003c\/h2\u003e\n\u003cp\u003e\u003cstrong\u003e1.Transportation conditions\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp\u003eThe whole process of low-temperature ice box transportation, to ensure that the kit is in a state of \u0026lt;4 °C.\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003e2. Storage conditions\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp\u003eThe kit is stored at -20℃. Store the product in a constant temperature refrigerator at -20℃ immediately after receipt. If the storage conditions are appropriate, the product will not degrade any performance during the 1-year validity period.\u003cbr\u003e2×RT-qPCR EasyTM Mix-SYBR component contains SYBR Green I, please keep it away from light.\u003c\/p\u003e\n\u003ch2\u003eTemplate RNA concentration\u003cbr\u003e\n\u003c\/h2\u003e\n\u003cp\u003eRT-qPCR EasyTM (One Step)-SYBR Green I:(0.1pg-100ng total RNA)\/20μl system.\u003c\/p\u003e\n\u003ch2\u003eOperation guide\u003cbr\u003e\n\u003c\/h2\u003e\n\u003cp\u003e\u003cstrong\u003eA: Preparation of materials and reagents\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp\u003e1. Prepare the prepared RNA template (it is recommended to use Foregene Total RNA Isolation Kit series kits to extract and purify RNA), specific primers (10μM) and related consumables and instruments.\u003c\/p\u003e\n\u003cp\u003eNote: Please ensure the integrity of RNA and try to use RNA extracted from fresh samples.\u003c\/p\u003e\n\u003cp\u003e2. Put 2×RT-qPCR EasyTM Mix-SYBR, RNase-Free ddH2O, and 50×ROX Reference Dye (if necessary) on ice to let it melt naturally, and flick the tube wall to mix until use.\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eB:RT-qPCR system preparation\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp\u003e2× RT-qPCR EasyTM Mix-SYBR is convenient and quick to use, and avoids pollution during operation and experimental errors caused by multiple preparations of the reaction system to the greatest extent. When using, only half the volume of the reaction system (for example, if the reaction system is 20μl, take 10μl 2×RT-qPCR EasyTM Mix-SYBR solution), add RNA template and specific primers, and add RNase-Free ddH2O to make up the volume. Refer to Table 1 below for the specific RT-qPCR reaction system preparation\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eForm 1:RT-qPCR system preparation\u003c\/strong\u003e\u003c\/p\u003e\n\u003ctable border=\"1\" style=\"width: 90%; border-collapse: collapse;\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 33.3333%; text-align: center; vertical-align: middle;\"\u003eRT-qPCR system additions\u003c\/td\u003e\n\u003ctd style=\"width: 33.3333%; text-align: center; vertical-align: middle;\"\u003eAmount\u003c\/td\u003e\n\u003ctd style=\"width: 33.3333%; text-align: center; vertical-align: middle;\"\u003eFinal concentration\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 33.3333%; text-align: center; vertical-align: middle;\"\u003e2× RT-qPCR EasyTM Mix-SYBR\u003c\/td\u003e\n\u003ctd style=\"width: 33.3333%; text-align: center; vertical-align: middle;\"\u003e10μl\u003c\/td\u003e\n\u003ctd style=\"width: 33.3333%; text-align: center; vertical-align: middle;\"\u003e1×\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 33.3333%; text-align: center; vertical-align: middle;\"\u003eForward Primer (10μM)\u003c\/td\u003e\n\u003ctd style=\"width: 33.3333%; text-align: center; vertical-align: middle;\"\u003e0.5μl\u003c\/td\u003e\n\u003ctd style=\"width: 33.3333%; text-align: center; vertical-align: middle;\"\u003e0.2-0.25μM 1*\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 33.3333%; text-align: center; vertical-align: middle;\"\u003eReverse Primer (10μM)\u003c\/td\u003e\n\u003ctd style=\"width: 33.3333%; text-align: center; vertical-align: middle;\"\u003e0.5μl\u003c\/td\u003e\n\u003ctd style=\"width: 33.3333%; text-align: center; vertical-align: middle;\"\u003e0.2-0.25μM 1*\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 33.3333%; text-align: center; vertical-align: middle;\"\u003eTemplate(RNA)\u003c\/td\u003e\n\u003ctd style=\"width: 33.3333%; text-align: center; vertical-align: middle;\"\u003eXμl\u003c\/td\u003e\n\u003ctd style=\"width: 33.3333%; text-align: center; vertical-align: middle;\"\u003e0.1pg-100ng\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 33.3333%; text-align: center; vertical-align: middle;\"\u003e50× ROX Reference Dye\u003c\/td\u003e\n\u003ctd style=\"width: 33.3333%; text-align: center; vertical-align: middle;\"\u003e-\u003c\/td\u003e\n\u003ctd style=\"width: 33.3333%; text-align: center; vertical-align: middle;\"\u003e2*\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 33.3333%; text-align: center; vertical-align: middle;\"\u003eRNase-FreeddH2O\u003c\/td\u003e\n\u003ctd style=\"width: 33.3333%; text-align: center; vertical-align: middle;\"\u003e(9-X)μl\u003c\/td\u003e\n\u003ctd style=\"width: 33.3333%; text-align: center; vertical-align: middle;\"\u003e \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 33.3333%; text-align: center; vertical-align: middle;\"\u003eTotal Volume\u003c\/td\u003e\n\u003ctd style=\"width: 33.3333%; text-align: center; vertical-align: middle;\"\u003e20μl\u003c\/td\u003e\n\u003ctd style=\"width: 33.3333%; text-align: center; vertical-align: middle;\"\u003e \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003cp\u003e1: Usually the final concentration of primer is 0.2-0.25μM to get better results. When the reaction performance is poor, the primer concentration can be adjusted within the range of 0.1-0.5μM.\u003c\/p\u003e\n\u003cp\u003eNote: Forward Primer and Reverse Primer are specific primers for the target gene.\u003c\/p\u003e\n\u003cp\u003e2: Choose the appropriate final concentration of ROX Reference Dye according to different quantitative PCR instruments. The optimal concentration of ROX Reference Dye for common quantitative PCR machines is shown in the following table:\u003c\/p\u003e\n\u003ctable border=\"1\" style=\"width: 90%; border-collapse: collapse;\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 50%; text-align: center; vertical-align: middle;\"\u003eQuantitative PCR instrument\u003c\/td\u003e\n\u003ctd style=\"width: 50%; text-align: center; vertical-align: middle;\"\u003eROX Reference Dye final concentration\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 50%; text-align: center; vertical-align: middle;\"\u003eABI PRISM 7000\/7300\/7700\/7900HT\/Step One etc.\u003c\/td\u003e\n\u003ctd style=\"width: 50%; text-align: center; vertical-align: middle;\"\u003e5× (i.e.20μl system, add 2μl 50×ROX Reference Dye)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 50%; text-align: center; vertical-align: middle;\"\u003eABI 7500,7500 Fast, Stratagene Mx3000P, Mx3005P, and Mx4000, etc.\u003c\/td\u003e\n\u003ctd style=\"width: 50%; text-align: center; vertical-align: middle;\"\u003e1× (i.e.20μl system, add 0.4μl 50×ROX Reference Dye)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 50%; text-align: center; vertical-align: middle;\"\u003eRoche PCR instrument, Bio-Rad PCR instrument, Eppendorf quantitative PCR instrument, etc.\u003c\/td\u003e\n\u003ctd style=\"width: 50%; text-align: center; vertical-align: middle;\"\u003eNo need to add ROX Reference Dye\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eC:RT-qPCR reaction system setting\u003c\/strong\u003e\u003c\/p\u003e\n\u003cul\u003e\n\u003cli\u003e\n\u003cp\u003eAfter preparing the RT-qPCR system with reference to the above table, mix gently (you can use a pipette tip to gently pipette; you can also mix on a vortexer and centrifuge briefly to collect the liquid scattered on the tube wall or tube cap, and place on ice box for later use).\u003c\/p\u003e\n\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003e2. Refer to the RT-qPCR reaction program settings (Table 2) to set the temperature and time of the reaction.\u003c\/p\u003e\n\u003cp\u003eNote: In order to ensure the activity of 2×RT-qPCR EasyTM Mix-SYBR and improve its amplification efficiency, it is best to prepare the RT-qPCR reaction system after setting up the PCR instrument program, so that the reaction program can be entered immediately after the system preparation is completed .\u003c\/p\u003e\n\u003cp\u003e3. In order to get the best qPCR effect, gradient PCR can be used to optimize the reaction conditions for different templates and different primers.\u003c\/p\u003e\n\u003cp\u003eNote: The extension temperature range of Foregene Hotstar Taq DNA Polymerase provided in this kit is: 60-72℃, and the best extension temperature is 72℃.\u003c\/p\u003e\n\u003cp\u003eThe following is an example of RT-qPCR reaction conditions. It is recommended to use a two-step method for PCR reaction. When the template concentration is too low to cause non-specific amplification, low primer Tm value leads to low amplification efficiency or poor amplification curve repeatability, etc., it is recommended to try the three-step method for PCR reaction. Refer to Table 2-1 (two-step method) and Table 2-2 (three-step method) for the setting of RT-qPCR reaction conditions.\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eForm 2-1:RT-qPCR reaction program setting(two step)\u003c\/strong\u003e\u003c\/p\u003e\n\u003ctable style=\"width: 90%; border-collapse: collapse;\" border=\"1\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003eStep\u003c\/td\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003eTemperature\u003c\/td\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003eTime\u003c\/td\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003eCycles\u003c\/td\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003eContent\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003e1\u003c\/td\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003e42℃\u003c\/td\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003e10-30min 1*\u003c\/td\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003e1\u003c\/td\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003eReverse Transcription\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003e2\u003c\/td\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003e94-95℃\u003c\/td\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003e5min\u003c\/td\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003e1\u003c\/td\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003ePre-denaturation\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\" rowspan=\"2\"\u003e3\u003c\/td\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003e94-95℃\u003c\/td\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003e10sec\u003c\/td\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\" rowspan=\"2\"\u003e30-45\u003c\/td\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003eTemplate denaturation during cycles\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003e60-65℃\u003c\/td\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003e20sec 2*\u003c\/td\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003eAnnealing\/Extension\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eForm 2-2:RT-qPCR reaction program setting( three step)\u003c\/strong\u003e\u003c\/p\u003e\n\u003ctable border=\"1\" style=\"width: 90%; border-collapse: collapse;\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003eStep\u003c\/td\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003eTemperature\u003c\/td\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003eTime\u003c\/td\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003eCycles\u003c\/td\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003eContent\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003e1\u003c\/td\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003e42℃\u003c\/td\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003e10-30min 1*\u003c\/td\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003e1\u003c\/td\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003eReverse Transcription\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003e2\u003c\/td\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003e94-95℃\u003c\/td\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003e5min\u003c\/td\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003e1\u003c\/td\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003ePre-denaturation\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"3\" style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003e3\u003c\/td\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003e94-95℃\u003c\/td\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003e10sec\u003c\/td\u003e\n\u003ctd rowspan=\"3\" style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003e30-45\u003c\/td\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003eTemplate denaturation during cycles\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003e55-65℃\u003c\/td\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003e20sec\u003c\/td\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003ePrimer annealing\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003e72℃\u003c\/td\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003e20-30sec 2*\u003c\/td\u003e\n\u003ctd style=\"width: 20%; text-align: center; vertical-align: middle;\"\u003eExtension\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003cp\u003e1: Reverse transcription time can be adjusted according to experimental needs. General endogenous genes such as β-Actin only need 10 minutes; to detect specific expressed genes, reverse transcription time can be appropriately extended as needed.\u003c\/p\u003e\n\u003cp\u003e2: Set a specific time according to the length of the amplified target fragment. The amplification speed of Foregene HotStar Taq DNA Polymerase is 2kb\/min.\u003c\/p\u003e\n\u003cp\u003eNote: PCR reaction conditions vary depending on the structural conditions of the template, primers, etc. For RNA templates with complex secondary structures, it is recommended to use 42℃ for the first step of reverse transcription. In the specific operation, it is necessary to design the optimal reaction conditions according to the specific conditions such as the size of the target fragment, the base sequence of the amplified fragment and the GC content and length of the primer, including annealing temperature, extension time, etc.\u003c\/p\u003e\n\u003ch2\u003eWork Flow\u003c\/h2\u003e\n\u003cdiv style=\"text-align: start;\"\u003e\u003cimg src=\"https:\/\/cdn.shopify.com\/s\/files\/1\/0521\/5312\/2997\/files\/RT-qPCR_EasyTM_I_One_Step_-SYBR_Green_I_work_flow_600x600.jpg?v=1752817236\" style=\"margin-bottom: 16px; float: none;\"\u003e\u003c\/div\u003e\n\u003cdiv style=\"text-align: start;\"\u003e\n\u003ch2\u003ePrecautions: (Please read the precautions carefully before using the kit)\u003c\/h2\u003e\n\u003cul\u003e\n\u003cli\u003e\n\u003cp\u003eReagents should avoid repeated freezing and thawing, otherwise the performance of the reagents will decrease or become invalid.\u003c\/p\u003e\n\u003c\/li\u003e\n\u003cli\u003e\n\u003cp\u003eThe reagents in the kit should be protected from light and stored at -20°C.\u003c\/p\u003e\n\u003c\/li\u003e\n\u003cli\u003e\n\u003cp\u003eIt is recommended to use fresh sample extraction or template RNA stored at -80°C (RNA should avoid repeated freezing and thawing).\u003c\/p\u003e\n\u003c\/li\u003e\n\u003cli\u003e\n\u003cp\u003eIn order to avoid RNase contamination, please perform the experiment in the RNase-Free space; the pipette tips and PCR tubes used must be RNase-Free; and wear disposable gloves and masks.\u003c\/p\u003e\n\u003c\/li\u003e\n\u003cli\u003e\n\u003cp\u003eThis kit must be used with specific primers for experiments. Please select the specific primers for the gene to be amplified according to the needs of the experiment.\u003c\/p\u003e\n\u003c\/li\u003e\n\u003cli\u003e\n\u003cp\u003eBefore use, put 2×RT-qPCR EasyTM Mix-SYBR I on ice to completely melt, flick and mix well before use; the preparation of the system should be operated on an ice bath to improve the performance of the kit and increase PCR amplification The specificity.\u003c\/p\u003e\n\u003c\/li\u003e\n\u003cli\u003e\n\u003cp\u003e2×RT-qPCR EasyTM Mix-SYBR contains SYBR Green I, please avoid strong ligh\u003c\/p\u003e\n\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003ch2\u003eDocuments\u003c\/h2\u003e\n\u003cp\u003e\u003ca href=\"https:\/\/cdn.shopify.com\/s\/files\/1\/0521\/5312\/2997\/files\/RT-qPCR_Easy_I_One_Step_-SYBR_Green_I_Instruction_Manual-V1.0-EN.docx?v=1752836237\"\u003e\u003cstrong\u003eRT-qPCR Easy™ I (One Step)-SYBR Green I Instruction Manual-V1.0-EN\u003c\/strong\u003e\u003c\/a\u003e\u003c\/p\u003e\n\u003c\/div\u003e","brand":"Foregene","offers":[{"title":"100 rxns","offer_id":51362514993333,"sku":"RT-02111","price":536.0,"currency_code":"USD","in_stock":true},{"title":"500 rxns","offer_id":51362515026101,"sku":"RT-02112","price":2143.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0521\/5312\/2997\/files\/RT-02111RT-qPCREasyTMI_OneStep_-SYBRGreenI_1.jpg?v=1752815007","url":"https:\/\/biofargo.com\/products\/rt-qpcr-easytm-i-one-step-sybr-green-i","provider":"Biofargo","version":"1.0","type":"link"}