Two cdna synthesis primers are provided with the kit. Using superscript iii firststrand synthesis supermix. The product of the first strand cdna synthesis reaction can be used directly in pcr. This kit provides all components for cdna synthesis, including reverse transcriptase, rnase inhibitor, oligo dt, random hexamer, dntps and nuclease free water. Hiscript ii 1st strand cdna synthesis kit interchim. Kit includes high capacity reverse transcriptase 100ul, optimized 5x buffer, random hexaprimer 20m and oligo dt primer 20m. Allinone mirna firststrand cdna synthesis kit in combination with miprofile mirna qpcr arrays and allinone qpcr mix. The length of cdna achieved is verified by detection of a 9. Prepare the following cdna synthesis mix, add the following components in the indicated order. This cdna synthesis kit provides all required reagents, except rna template and genespecific primers, to create first strand cdna from an rna template.
Six replicate cdna reactions were performed for each input quantity of rna. Accuris qmax first strand cdna synthesis flex kit, sample pack, 5 reactions catalog number pr2110s. First strand cdna synthesis after thawing, mix and briefly centrifuge the components of the kit. The first strand cdna synthesis flex kit works optimally with 10pg to 2. First strand cdna synthesis section vii since the 5 elongation benefits of smart technology are only relevant for 5race, the smart race kit includes a protocol for the synthesis of two separate cdna populations.
Maxima first strand cdna synthesis kit for rtqpcr, with. The accuris qmax first strand cdna synthesis flex kit is available in 50 and 200 reaction packs. Product size sku vitascript firststrand cdna synthesis kit 50 rxn 20 l pccsku1 component volume 5x vs reaction buffer 4 l vitascript enzyme mix 1 l total rna 16 l nucleasefree dh 2. K1612, first strand cdna synthesis, rev15 thermo fisher scientific. The sensitivity of the kit is verified by the detection of gapdh transcript in 20 pg total rna after 35 cycles. Mmlv reverse transcriptase 1ststrand cdna synthesis kit lucigen. First strand cdna synthesis kit 11801100 delivery time may vary from web posted schedule. It provides a dramatically simplified workflow that combines genomic dna elimination and cdna synthesis into onetube procedure. For following pcr applications, the diluted cdna reaction should represent 10% of the total reaction volume e. Reverse transcriptase, which has lower rnase h activity compared to amw reverse transcriptase. The vazyme hiscript ii reverse transcriptase is a new generation reverse transcriptase optimized from.
The resulting singlestranded cdna is suitable for use in real time quantitative reverse transcription pcr rtqpcr. The mmlv reverse transcriptase 1ststrand cdna synthesis kit is optimized for generating fulllength firststrand cdna from total cellular rna or polya. The synthesis reaction can be completed in 15 to 30 minutes. The kit has been extensively tested with a variety of rna templates, including gc and aurich targets and rnas expressed at low levels.
The latter is designed to bind at the beginning of the polya tail to generate fulllength. First strand cdna synthesis employs the clontech smarter pcr cdna synthesis kit. Firststrand cdna synthesis kit for realtime pcr product code pack size 75780 50 reactions 20 l usb firststrand cdna synthesis kit for realtime pcr fig. The rt 2 first strand kit provides a rapid and convenient procedure for efficient first strand cdna synthesis and genomic dna elimination in rna samples. Leukemia virus mmlv reverse transcriptase to synthesize first strand cdna from. For priming, three different primer systems can be used. It can be used with either sequencespecific primers, polydt15 primers, or random primers, pdn6. Use 1 g of isolated mrna to generate first strand cdna for second strand synthesis and subsequent cloning reactions. Thermo scientific first strand cdna synthesis kit is a complete system for efficient synthesis of first strand cdna from mrna or total rna templates. Truscript first strand cdna synthesis kit for mrna product. The quality of the mrna preparation, the purity of the enzyme and reagents used in the first strand reaction, and the choice of second strand method affect the quantity. For random hexamer primed synthesis, incubate for 60 min at 37c. First strand cdna synthesis kit for rtpcr amv sigma.
Firststrand cdna synthesis kit introduction the firststrand cdna synthesis kit is based on reverse transcriptase cat. The length of cdna achieved is verified by the detection of a 5. For the first strand synthesis of singlestranded cdna from rna for use as a pcr template. The first strand cdna synthesis kit is used for the synthesis of the first strand cdna as the starting reaction for twostep rt pcr. Normally, 2 l of the first strand cdna synthesis reaction mixture is used as template for subsequent pcr in a 25 l total volume. Usb firststrand cdna synthesis kit for realtime pcr. Amrt0020 20 rt reactions, amrt0060 60 rt reactions used in combination with miprofile mirna qpcr arrays and allinone qpcr mix. The kit includes reverse transcriptase amv for first strand synthesis, two different primers, our pcr nucleotide mix, and control neo pa rna. The iscript cdna synthesis kit is a sensitive and easytouse first strand cdna synthesis kit for gene expression analysis using realtime qpcr. The iscript cdna synthesis kit is a sensitive and easytouse firststrand cdna synthesis kit for gene expression analysis using realtime qpcr.
Hyperscripttm first strand synthesis kit is ideally organized for synthesizing reaction of first strand cdna from purified mrna or total rna. Asfs002 product summary product description the rtstar first strand cdna synthesis kit 3 adaptor is designed to create cdna libraries from small rnas for qpcr detection. Nzy firststrand cdna synthesis kit is formulated to provide high yields of fulllength cdna products and to increase. Firststrand cdna synthesis quantabio reverse transcription reagents leverage proprietary, performanceengineered qscript reverse transcriptase in a variety of stabilized, userfriendly reagent formulations that maximize cdna yield and provide linear cdna synthesis across a broad dynamic range of input rna. First strand cdna synthesis was carried out with 1x mmulv enzyme mix at 42c using 2 g of human spleen total rna. Increase the temperature of first strand reaction up to 55c. The first strand cdna synthesis kit is used for the synthesis of the first strand cdna as the starting reaction for rtpcr. Aomdq020 20 rt and 200 qpcr reactions, aomdq050 50 rt and. June 2017 the transcriptor first strand cdna synthesis kit is designed for first strand cdna synthesis reactions up to 14 kb. The cdna synthesis system provides the materials needed to rapidly and reliably.
Hyperscript first strand synthesis kit 1 bohemia genetics. The products can be used for pcr immediately or be stored at 20. Nzy mmulv firststrand cdna synthesis kit, separate oligos. This has been shown to increase sensitivity in qrtpcr. The kit uses mmulv vity compared to amw reverse transcriptase. Select cdna synthesis kit iscript biorad laboratories. Maxima first strand cdna synthesis kit for rtqpcr, with dsdnase.
This variation can enter in any of the three steps of cdna synthesis. The amv enzyme mix is an optimized blend of amv reverse transcriptase and a murine rnase inhibitor. Nzy firststrand cdna synthesis kit cdna kits nzytech. Transcriptor first strand cdna synthesis kit uses anchored oligodt priming and with this kit, the complete. The mirx mirna first strand synthesis kit included in the mirx mirna qrtpcr tb green kit and also sold separately is used for converting mirnas, and other rna molecules, into cdna to enable specific rnas to be quantified by realtime pcr figure 1. The volume of the first strand cdna synthesis reaction mixture should not comprise more than 110 of the total pcr reaction volume. Truscript first strand cdna synthesis kit for mrna product insert. First strand cdna synthesis protocols e6300 introduction. Description biotechrabbit cdna synthesis kit provides superior components that ensure efficient firststrand. However, this step can be omitted in some cases unpublished results. Quantitative first strand synthesis of all rna species is achieved with the use of the random primers. The kit uses revertaid reverse transcriptase rt, a recombinant mmulv rt which maintains activity at 4250c and is suitable for synthesis of cdna up to kb. First strand cdna synthesis kit, separate oligos, is formulated to provide high yields of fulllength cdna products and to increase sensitivity in rtqpcr. This twotube kit is optimized to yield sensitive, unbiased representation over a broad dynamic range, with minimal setup.
The transcriptor first strand cdna synthesis kit is designed to reverse transcribe rna mrna, total rna, viral rna, and in vitro transcribed rna from a. Primers synthesis of first strand cdna can be primed with either oligodt 18 primer, random primers or genespecific primers. The kit provides all reagents required for first strand cdna synthesis reactions. Oligodt 15 primer vial 5 for standard cdna synthesis followed by cloning or restriction enzyme. Maintain a separate set of automatic pipettes for manipulating rna. Thermo scientific maxima first strand cdna synthesis kit for rtqpcr with dsdnase is a convenient system optimized for cdna synthesis in 2step quantitative rtpcr rtqpcr applications. Amv reverse transcriptase provides first strand cdna synthesis reactions with a broader optimal reaction temperature from 37c to 50c. The synthesized cdna is ready to use in realtime pcr expression analysis of multiple genes. Accuscript high fidelity 1st strand cdna synthesis kit. This kit allows reverse transcription at high temperatures and without rnase h activity, ensuring efficient reverse transcription of targets with strong secondary structures and providing increased specificity, higher yields of cdna. This supermix formulation can be used to quantify fewer than 10 copies of a target gene in qrtpcr, with a broad dynamic range that supports. A control reaction without reverse transcriptase is recommended to examine the dna contamination in the samples.
Reverse transcriptase, which has lower rnase h acti first. As it is well known, trnas undergo by far the greatest number of and the most chemically diverse posttranscriptional modifications, which hinder the cdna synthesis. High quality, pcr ready cdna is quickly generated with the accuris qmax first. A genetically engineered version of mmlv reverse transcriptase with no rnase h activity is a rnadirected dna enzyme that synthesizes a complementary dna strand initiating from a primer using single. In the beginning, the reverse transcriptase enzyme amplifies the cdna from the singlestranded rna, known as a first strand cdna synthesis. First strand cdna synthesis with high sensitivity and efficiency. The cdna synthesis is completed into a twostep, first strand cdna synthesis followed by second strand dna synthesis. Recommended quantities of primer for first strand synthesis in 1. Truscript first strand cdna synthesis kit product insert. This kit produces an optimal template for dna amplification techniques. Components of the maxima first strand cdna synthesis kit for rtqpcr are premixed to save. The synthesis of cdna is a crucial step in the entire rtpcr protocol. Protoscript first strand cdna synthesis kit e6300 manual neb.
Protoscript ii first strand cdna synthesis kit e6560 manual. Amv reverse transcritase is used for first strand cdna synthesis. When performing reverse transcription qpcr and cdna synthesis for qpcr, you need reagents that deliver high specificity and sensitivity and process safety. Using the fermentas first strand cdna synthesis kit quantify we need 0. Power cdna synthesis kit is used to synthesize firststrand cdna from total rna or mrna template. It contains all components necessary for cdna synthesis. 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.
Synthesis kit is a complete system for efficient synthesis of first strand cdna from mrna or total rna templates. Thermo scientific revertaid first strand cdna synthesis kit is a complete system for efficient synthesis of first strand cdna from rna templates. We recommend incubation at 42c for one hour for maximum yield and length. Transcriptor first strand cdna synthesis kit y version.
The resulting singlestranded cdna is suitable for use in realtime quantitative reverse transcription pcr rtqpcr. Our rtqpcr kits have a unique combination of inprocess controls including a builtin visual indicator to remove variables and increase the accuracy, specificity and sensitivity of your analyses. Thaw 5x paprt buffer and ddh2o rnase dnase free at room temperature 1525c. The performance of protoscript first strand cdna synthesis kit is tested in an rt reaction using human jurkat total rna with primer dt 23vn. The first strand cdna plus synthesis kit contains all reagents for first strand cdna synthesis and offers a simple handling with high flexibility.
Assay sensitivity and dynamic range various amounts of hela cell total rna, ranging from 850 ng down to 10 pg, were reverse transcribed using anchored oligodt 23 vn as. The performance of protoscript ii first strand cdna synthesis kit is tested in an rt reaction using jurkat total rna with primer dt 23vn. Superscript iii firststrand synthesis supermix for qrtpcr. Reverse transcriptase is a new enzyme obtained through genetic engineering based on mmlv rnase h reverse transcriptase. First strand cdna synthesis kit for rtpcr amv sigmaaldrich. Occasional delays may occur due to unforeseen complexities in the preparation of your product. Mix rna sample and primer dt 23 vn in two sterile rnasefree microfuge tubes. Superscript iii firststrand synthesis system for rtpcr. This twotube kit is optimized to yield sensitive, unbiased representation over a broad dynamic range, with minimal setup and reaction time. Either the transcriptor first strand cdna synthesis kit blue curves or another supplierss reverse transcriptase red curves was used to perform reverse transcription on 100 ng to 10 pg of total rna from k562 cells. Nzy firststrand cdna synthesis kit is a system that includes all the necessary components to synthesize first strand cdna, except the template rna. Protoscript first strand cdna synthesis kit e6300 manual.
Lightcycler faststart dna masterplus sybr green i, was then used with primers for g6pdh to quantify geneexpression levels. The amv first strand cdna synthesis kit combines amv enzyme mix and amv reaction mix. The reaction product of the first strand cdna synthesis can be used directly in qpcr, or stored at 20 c for up to one week. First strand cdna synthesis reaction denaturation of rna and primer at 70c for 5 minutes can remove secondary structures that may impede long cdna synthesis. Not recommended or intended for diagnosis of disease in humans or animals. All kit components are optimized to facilitate efficient cdna synthesis using 1 pg to 1 g of input total rna. Accuscript high fidelity 1st strand cdna synthesis kit agilent. The transcriptor high fidelity cdna synthesis kit is designed to reverse transcribe rna with an increased fidelity cat.
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