{"product_id":"rapidxfire-thermostable-reverse-transcriptase-307","title":"RapiDxFire Thermostable Reverse Transcriptase","description":"\u003cp\u003eRapiDxFire Thermostable Reverse Transcriptase is a significant advancement over popular MMLV- and AMV reverse transcriptases. Originally identified in hot springs, this robust bacteriophage enzyme exhibits increasingly high activity at elevated temperatures (~60% activity remains after 10 min at 90 ⁰C) and maintains its core activity after \u0026gt;3 months storage at 20-25⁰C. This enzyme lacks RNAse H and 3’ 5’ exonuclease activity and efficiently synthesises short cDNA fragments (≤ 1 Kb). Like other reverse transcriptases, RapiDxFire RT possesses DNA polymerase activity but lacks 5′ 3′ exonuclease activity. Supplied in a glycerol-free, Triton™ X-100-free storage buffer, the enzyme formulation can be further optimised for downstream lyophilisation.\u003c\/p\u003e\u003cp\u003eRapiDxFire Thermostable RT Performs Best at High Temperatures\u003c\/p\u003e\u003cp\u003eFigure 1. Reaction temperature profile for RapiDxFire Thermostable Reverse Transcriptase. Reverse transcription reactions were set up on ice using each manufacturer’s recommended buffer system and Poly (rC) \/p(dG)12-18 template\/primer substrate. Reactions were transferred from ice to the indicated temperatures (37, 50, 55, 60, 65, 70, 75, and 80 °C) and incubated for 40 minutes. Following incubation, RNA\/cDNA heteroduplex product is quantified as a measure of polymerisation activity, utilising PicoGreen® fluorescence on a Tecan Infinite® M1000 Pro. RapiDxFire Thermostable RT exhibits increasing activity as the reaction temperature is increased, up to 80 °C (highest temperature tested); ~60% activity remains after 10 min at 90 °C (data not shown).\u003c\/p\u003e\u003cp\u003eSuperior Detection of Zika virus with RapiDxFire Thermostable RT\u003c\/p\u003e\u003cp\u003eFigure 2. cDNA synthesis time course studies in a 2-step RT-qPCR reaction with different thermostable RTs. A) qPCR curve after a 1 minute reverse transcription reaction. Zika cDNA synthesis was conducted for each enzyme in duplicate using target-specific primers and recommended reaction buffer and incubation temperatures per suppliers’ guidelines (RapiDxFire= 60 ⁰C, Supplier B= 50 ⁰C, and Supplier T= 55 ⁰C). After cDNA synthesis real time PCR was performed using one-tenth volume of each of the cDNA samples. PCR was performed using EconoTaq and its supplied buffer (Item No. 30031-3). Detection of PCR products were done real time using an intercalating dye (Dyomics Cat No. V13-01184) and BioRad CFX C1000 Touch™ with absorption\/emission of 481nm\/526nm. B) Reverse transcription time course study prior to performing second-step real-time PCR. Encircled data points are derived from data represented in the qPCR curve in A).\u003c\/p\u003e\u003cp\u003e3-month Room Temperature Stability\u003c\/p\u003e\u003cp\u003eFigure 3. RapiDxFire Thermostable RT was stored in separate aliquots per time point at ambient and -20 °C. At each time point RapiDxFire Thermostable RT was evaluated by measuring first-strand cDNA synthesis of 10000 copies MS2 RNA. cDNA synthesis was carried out using a gene-specific primer for MS2 at reaction temperature of 60 °C for 5 minutes. cDNA was measured using real time PCR and MS2 primers design around a 520 base pair amplicon.\u003c\/p\u003e\u003cp\u003eLegal Information\/Updates:\u003cbr\u003eUS Patent 8093030 for RapiDxFire™ is owned by LGC, Biosearch Technologies. For research use only. Not for use in diagnostic procedures.\u003c\/p\u003e\u003cp\u003eread more\u003c\/p\u003e\n\u003ch2\u003eKey features\u003c\/h2\u003e\n\u003cul\u003e\n\u003cli\u003eExtremely active at high temperatures (55 °C to 80 °C): Improves specificity of cDNA Synthesis from diverse RNA templates\u003c\/li\u003e\n\u003cli\u003eSensitive: Detects 100 copies of RNA in two-step RT-qPCR assays\u003c\/li\u003e\n\u003cli\u003eShort reaction time (5 minutes or less): Streamlined RT-qPCR workflow and faster time to results\u003c\/li\u003e\n\u003cli\u003eStable at room temperature (\u0026gt; 3 months): Simplifies setup on automation decks (no cold storage required) and enables use in environments with limited cold-storage\u003c\/li\u003e\n\u003cli\u003eLyo-compatible: Enzyme formulation is free of glycerol -and other components- that are known to interfere with downstream lyophilisation\u003c\/li\u003e\n\u003cli\u003eBatch to batch reproducibility: Manufactured in an ISO 13485-certified facility\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003ch2\u003eSpecifications\u003c\/h2\u003e\n\u003cp\u003eStorage: Store at -20 °C.\u003c\/p\u003e\u003cp\u003eStorage Buffer: RapiDxFire Thermostable RT is supplied in a glycerol-free, Triton X-100-free buffer.\u003c\/p\u003e\u003cp\u003eUnit Definition: One RT unit is defined as the amount of enzyme required to incorporate 1 nmol of dTTP into an acid-insoluble form in 10 minutes at 55 °C (using poly(rA) oligo(dT) as template primer and 1X kit -supplied buffer formulation).\u003c\/p\u003e\u003cp\u003eFunctional QC Assay: RapiDxFire Thermostable RT is functionally tested with gene-° primer and bacteriophage MS2 RNA. In this reaction the enzyme converts 1000 copies of MS2 RNA into 520 bp cDNA in 5 minutes at 60 °C.\u003c\/p\u003e\u003cp\u003eEndonuclease: Free of contaminating endonucleases at the 10 Unit level.\u003c\/p\u003e\u003cp\u003eExonuclease: Free of contaminating exonucleases at the 10 Unit level.\u003c\/p\u003e\u003cp\u003eRNase Activity: No RNase activity detected using a fluorescence-based RNase activity assay.\u003c\/p\u003e\u003cp\u003eread more\u003c\/p\u003e\n\u003ch2\u003eProduct Details\u003c\/h2\u003e\n\u003cp\u003eContents include RapiDxFire Thermostable Reverse Transcriptase (3 U\/µL) and 10X RapiDxFire Reaction Buffer. The enzyme pack sizes (50 or 250 Rxns) are based on 25 µL reaction volumes.\u003c\/p\u003e\n\u003ch2\u003eResources\u003c\/h2\u003e\n\u003cp\u003eBrochures and guides\u003c\/p\u003e\u003cp\u003eapp note rapidxfire reverse transcriptase\u003c\/p\u003e\n\u003ch2\u003eDocuments\u003c\/h2\u003e\n\u003cul\u003e\n\u003cli\u003e\u003ca href=\"https:\/\/biosearchtech.a.bigcontent.io\/v1\/static\/sds_307_30250-RapidxFire-Thermostable-RT\"\u003e30250 RapidxFire Thermostable RT\u003c\/a\u003e\u003c\/li\u003e\n\u003cli\u003e\u003ca href=\"https:\/\/biosearchtech.a.bigcontent.io\/v1\/static\/manual_307_RapiDxFire-reverse-transcriptase-manual\"\u003eRapiDxFire reverse transcriptase manual\u003c\/a\u003e\u003c\/li\u003e\n\u003cli\u003e\u003ca href=\"https:\/\/biosearchtech.a.bigcontent.io\/v1\/static\/guide_307_app-note-rapidxfire-reverse-transcriptase\"\u003eapp note rapidxfire reverse transcriptase\u003c\/a\u003e\u003c\/li\u003e\n\u003c\/ul\u003e","brand":"Biosearch Technologies","offers":[{"title":"50 x 25 µL Reactions","offer_id":45986431467599,"sku":"30250-1","price":2770.0,"currency_code":"ILS","in_stock":true},{"title":"250 x 25 µL Reactions","offer_id":45986431500367,"sku":"30250-2","price":0.0,"currency_code":"ILS","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0712\/0748\/5519\/files\/RapiDxFire-Thermostable-Reverse-Transcriptase_cbcc5f5c-0cfb-465a-ac15-09668a324b7d.jpg?v=1785956113","url":"https:\/\/www.danyel.co.il\/products\/rapidxfire-thermostable-reverse-transcriptase-307","provider":"danyel biotech","version":"1.0","type":"link"}