Protocol for cdna synthesis and qrtpcr cdna synthesis. Highcapacity cdna reverse transcription kits catalog numbers 43688, 4368814, 4374966, and 4374967 pub. I want to synthesize cdna for qpcr using the iscript cdna synthesis kit. The random primer method is used for initiating cdna synthesis, and ensure that first strand synthesis.
Rna isolationpurification and firststrand cdna synthesis. Choose the best cdna synthesis kit for your application, with formulations optimized for ease, yield, flexibility, fidelity, or gdna clearance. The qscript flex cdna synthesis kit is an easytouse and highly efficient kit for the synthesis of firststrand cdna that enables your choice of cdna priming method. The sensifast cdna synthesis kit has been designed for rapid generation and unbiased coverage of cdna, especially for subsequent quantification with realtime pcr.
The qiagen rneasy mini kit 2 is useful in removing contaminants and impurities from an rna sample isolated from other methods. Rnase and s1 nuclease protection cdna synthesis differential display expressionarray and expressionchip analysis in addition, rneasy kits can be used to desalt or purify rna from enzymatic. The enzyme maintains activity at 37c and is suitable for synthesis of cdna up to 9 kb. Find the qiagen plasmid kit to suit your exact needs using the new online selection. The kit utilizes our smart switching mechanism at 5 end of rna template technology coupled with pcr amplification to generate high yields of fulllength doublestranded cdna suitable for various applications, including clontech pcrselect cdna subtraction, nondirectional cloning, and preparation of. It is important to have intact rna as starting template. Toyobo has various kits for cdna synthesis based on the genetically improved mmlv reverse transcriptase revertra ace. The rnaquant cdna synthesis kit prepares any rna for qpcr. Products cdna synthesis kits toyobo biotech support. Rna isolation from embryonic zebrafish and cdna synthesis. The kit uses mmulv vity compared to amw reverse transcriptase.
No amplification was observed from biorad iscript gdna clear cdna synthesis kit a, demonstrating complete removal of gdna. Paxgene blood mirna kit brochure qiagen pdf catalogs. Thermo scientific first strand cdna synthesis kit is a complete system for efficient synthesis of first strand cdna from mrna or total rna templates. Synthesis of highly structured and long cdna fragments, extremely sensitive and highly specific rtpcr, dna labeling. For cdna synthesis with integrated removal of genomic dna. By use of this product, you accept the terms and conditions outlined in the licensing and warranty statement containedinthis user manual. Script cdna synthesis kit contains all reagents required for first strand cdna synthesis in one box combining simple handling with high flexibility. Rna isolation from embryonic zebrafish and cdna synthesis for. Some are designed strictly for firststrand cdna synthesis. For cdna synthesis with integrated removal of genomic. Mirx uses an optimized mix of polya polymerase and smart mmlv reverse transcriptase to synthesize firststrand cdna.
Efficiency kits varied in their ability to efficiently generate cdna from different rna inputs linearity of cdna synthesis kits. Then, readyscript reverse transcriptase and other necessary reagents for cdna synthesis are subsequently added to convert the polya tailed micrornas into cdna using an oligodt adapter primer. As an alternative, double stranded cdna synthesis can be performed, instead of the first strand synthesis demonstrated in this protocol. The rt2 preamp cdna synthesis kit and rt2 preamp pathway primer mix are. Did anyone tried to see expression levels of microrna using cdna synthesis kit other than qiagen. Currently, all the mirna cdna synthesis kits on the market were cre.
Find product specific information including cas, msds, protocols and references. Steps to achieve quantitative measurements of microrna. Thermo scientific verso cdna synthesis kit provides robust transcription of rna to create a complete cdna pool. Total rna and mirna can also be copurified from formalinfixed, paraffinembedded.
January 2017 for the synthesis of doublestranded cdna from total rna or mrna cat. Fully optimized for realtime quantitative pcr qpcr applications, the kit utilizes our qpcrgrade affinityscript multiple temperature reverse transcriptase, including a buffer system optimized for realtime qpcr. Perfecting pcr is the goal quantabio has enthusiastically strived for in designing pcr and cdna synthesis reagents that define the standard for reproducibility, specificity and sensitivity. The taqman advanced mirna cdna synthesis kit uses 3 polya tailing and 5 ligation of an adaptor sequence to extend the mature mirnas present in the sample on each end prior to reverse. Reactions can be scaled up to 100 l to generate 10 g of cdna from a single reaction.
Applied biosystems high capacity cdna reverse transcription kit biorad iscript cdna synthesis kit invitrogen superscript iii firststrand synthesis supermix quanta qscript cdna synthesis kit quanta qscript cdna supermix qiagen quantitect reverse transcription kit stratagene affinityscript qpcr cdna synthesis kit. Sensifast cdna synthesis kit and a kit from supplier b were used for reverse transcription of a specific target from total rna under fast conditions. A single cdna synthesis reaction can be used for detection of hundreds of mirnas. Dna fragments yes yes yes yes oligonucleotides no yes no no ligation yes yes yes yes nick translation yes yes yes yes pcr yes no no yes random priming yes yes yes yes. For reverse transcription of 50 ng to 2 g rna per reaction for endpoint pcr. The results illustrate that the sensifast cdna synthesis kit red is more sensitive than the kit from supplier b blue, as judged by the lower ct values. May 12, 2009 qiagen s quantitect reverse transcription kit is designed to generate high quality cdna, which can be used for highly sensitive downstream applications such as realtime quantitative pcr qpcr. Using the fermentas first strand cdna synthesis kit quantify we need 0. The taqman advanced mirna cdna synthesis kit uses 3 polya tailing and 5 ligation of an adaptor sequence to extend the mature mirnas present in the sample on each end prior to reverse transcription. For cdna synthesis with integrated removal of genomic dna contamination for use in realtime twostep rtpcr. This kit includes the miscript universal primer which allows detection of. The kit uses mmlv reverse transcriptase with lower rnase h activity, which avoids degradation of rna when rnadna hybrid forms in the progress of cdna synthesis.
The choice of the adapter pair to use for a particular cdna preparation depends on intended application. The qscript cdna synthesis kit is a sensitive and easytouse solution for rna. The dual buffer system meets the distinctive needs of mirna quantification using realtime pcr. Highcapacity cdna reverse transcription kits product. From solutions from gels qiaquick qiaquick pcr nucleotide qiaquick gel qiaquick gel purification kit removal kit extraction kit extraction kit alkaline phosphatase yes yes yes yes cdna synthesis. Microrna firststrand synthesis and mirna quantitation kits. To be used in conjunction with the paxgene blood rna tubes paxgene blood rna tubes 100 100 blood collection tubes. In accordance with qiagen s isocertified quality management system, each lot of the rneasy mini kit, rneasy protect mini kit, and rneasy plant mini kit is tested against predetermined specifications to. The qscript cdna synthesis kit is a sensitive and easytouse solution for rna quantification using twostep rtpcr. Quantitect reverse transcription kit for cdna synthesis from. The novel qscript reaction mix provides all the necessary components for cdna synthesis except enzyme and rna template. En quantitect reverse transcription handbook qiagen. Rtpcr reverse transcription pcr reverse transcriptases.
The smarter pcr cdna synthesis kit is an improved version of our original smart pcr cdna synthesis kit. First strand cdna synthesis kit for rtpcr amv sigmaaldrich. Affinityscript qpcr cdna synthesis kit provides a streamlined master mix format and fast protocol, with a 15minute cdna synthesis step and an overall protocol time of just 25 minutes. The efficiency of the reaction is highly dependent on the quality and quantity of the starting rna template. Researchers can choose from among a wide variety of commercial cdna synthesis kits. The verso system achieves robust and sensitive reverse transcription through the. The novel qscript reaction mix provides all the necessary components for cdna synthesis. Mar 07, 2011 a complete system for highly efficient synthesis of first strand cdna. Unlike other vendors kits, these microrna quantitation kits provide a simple, singlestep protocol for the firststrand synthesis reaction. With the qscript microrna cdna synthesis kit mirnas are polyadenylated in a polya polymerase. The quantitect reverse transcription kit is shipped on dry ice. The high capacity cdna reverse transcription kit delivers extremely highquality, singlestranded cdna from 0. This kit enables simple, singlestep cdna synthesis. For purification of up to 10 mg transfectiongrade plasmid or cosmid dna.
Omniscript reverse transcriptase handbook 0499 9 protocol omniscript protocol for reverse transcription this is the standard protocol for firststrand cdna synthesis using 50 ng to 2 g rna and omniscript reverse transcriptase from qiagen. For highthroughput purification in a 96well format, the mirneasy 96 kit is available. For isolation of genomic, mitochondrial, bacterial. Show full abstract mirneasy mini kit qiagen according to the manufacturers instructions. Quantitect reverse transcription kit qiagen online shop. For highspeed preparation of cdna without rna purification for use in realtime rtpcr. The smarter pcr cdna synthesis kit provides a pcrbased method for producing highquality cdna from nanogram quantities of total rna. Components of this kit are used with the rt primer provided with the taqman microrna assay to convert mirna to cdna. The qscript microrna cdna synthesis kit has been designed to convert mirnas into cdna starting from total rna or rna preparations preenriched for mirnas. Qiagen markets this kit specifically for qpcr which. These are available online in convenient and compact pdf format at. Thermo scientific reverse transcription thermo fisher. Qiaquick kits can be used to cleanup the following enzymatic reactions. The thermo scientific revertaid first strand cdna synthesis kit is a complete system for efficient synthesis of first strand cdna from mrna or total rna templates.
Reverse transcription rt is a widely used application in molecular biology research for rna and expression analysis. High cdna yields due to highaffinity enzyme sensitive detection of as few as 10 copies of template no additional rnase h digestion step required fast and easy procedure with no tedious pipetting steps the omniscript reverse transcription rt kit. Reverse transcriptases are used in vitro for firststrand cdna synthesis with rna as the starting template. Please refer to the section below to choose the pair of adapters suitable for your needs. The tonkbio first chain cdna synthesis kit for rtpcr is optimized to efficiently synthesize first chain cdna from mrna or total rna templates. Purification kit removal kit extraction kit extraction kit alkaline phosphatase yes yes yes yes cdna synthesis yes no no yes dnase, yes yes yes yes nuclease digestion kinase. Lysis and reverse transcription are performed in the same tube, and the resulting firststrand cdna.
Affinityscript multiple temperature cdna synthesis kit achieved successful reads at the 0. Rnaquant cdna synthesis kit profile any rna by realtime qpcr rnaquant. The omniscript rt kit is intended for molecular biology applications. Universal rt primers recognize the universal sequences present on both the 5 and 3 extended ends of the mature mirnas. The superscript iii cellsdirect cdna synthesis kit is optimized for synthesizing firststrand cdna directly from a mammalian cell lysate without first isolating the rna. The reagents and kits included in this protocol have worked well and routinely yielded high quality rna in our laboratory.
Quantinova reverse transcription kit handbook qiagen. Sigmaaldrich offers roche11483188001, first strand cdna synthesis kit for rtreverse transcriptasepcr amv for your research needs. Mint2 cdna synthesis kit contains different plugoligo and cds adapters allowing synthesis of cdna with different flanking sequences. We are doing the rna extraction with the qiagen rneasy plant mini kit. Complementary dna compound, affinityscript cdna synthesis kit. Reverse transcriptase for cdnasynthesis efficiency and functional absence of rnases. Quanta qscript cdna synthesis kit quanta qscript cdna supermix qiagen quantitect reverse transcription kit stratagene affinityscript qpcr cdna synthesis kit. Learn about the affinityscript qpcr cdna synthesis kit, which offers the highest efficiency of conversion of rna to cdna. At just minutes, the lunascript rt supermix kit offers the shortest available firststrand cdna synthesis protocol comparison of recommended protocols for cdna synthesis. The qiagen onestep rtpcr kit is shipped on dry ice and should be stored immediately upon receipt at. For cdna synthesis and genomic dna elimination in 96 rna samples for use with rt 2 profiler pcr arrays. Reverse transcriptases rts use an rna template and a primer complementary to the rna to direct the synthesis of the first strand cdna, which can be used directly as a template for the polymerase chain reaction pcr.
Qiagen kit handbooks and user manuals are available at. Store kit at 20c on receipt a limiteduse label license covers this product. The thermo scientific reverse transcription portfolio offers robust and reliable. Downstream applications include realtime pcr, standard pcr, and microarrays. These are available online in convenient and compact pdf. Choose from a range of reverse transcriptases and rtpcr kits for reverse transcription of fulllength cdna from your experimental sample, including goscript reverse transcriptase, which uses mmlv reverse transcriptase and stateoftheart buffer to drive robust, reliable cdna synthesis of a full range of rare and abundant transcripts. The polya tailing and tagging method in the rnaquant system generates cdna that is highly unique and, when used with the 3 universal reverse primer, does not produce any signal in the absence of reverse transcriptase or lead to the detection of any residual genomic dna. The optimized 5x transamp buffer includes all necessary components for cdna synthesis. Pdf total rna isolation and cdna synthesis from bixa. If i used miscript ll qiagen, do i have to use sybr green kit qiagen. The iscript cdna synthesis kit performs across a broad range of concentrations. The lunascript rt supermix kit requires the shortest reaction time and tolerates elevated temperatures, reducing complications from rna secondary structure. Reverse transcriptase, which has lower rnase h acti first.
Choose the best cdna synthesis kit for your application, with. With the mysticq microrna cdna synthesis kit, micrornas are polyadenylated through a polya polymerase reaction. The extracted rna was successfully converted into doublestranded cdna using the smater cdna synthesis kit clontech, usa which is based on the switching mechanism at 5 end of rna transcript. Script cdna synthesis kit, reverse transcriptases with.