Nrevertaid first strand cdna synthesis kit pdf merger

This kit is powered by primescript reverse transcriptase primescript rt, which has exceptionally strong strand displacement activity. Avoid repeated freezethaw cycles for control neo pa rna, rnase. Quickly merge multiple pdf files or part of them into a single one. Accuscript high fidelity 1st strand cdna synthesis kit. The cdna synthesis procedure is illustrated in fig. In this experiment, cdnas prepared using each method were amplified in real time with primers specific for the 5 end of the adar gene using three different. Sensifast cdna synthesis kit, 50 reactions, first strand synthesis, fulllength reverse transcription. Protoscript ii first strand cdna synthesis kit features two optimized mixes, protoscript ii enzyme mix and protoscript ii reaction mix. Mmlv reverse transcriptase 1ststrand cdna synthesis kit lucigen.

Thermo scientific revertaid first strand cdna synthesis kit. The accupower rt premix is a master mix for cdna synthesis that consists of an easy to resuspend, lyophilized mix of mmlv reverse transcriptase, rnase inhibitor, dntps, reaction buffer, tracking dye, and patented stabilizer. The kit is highly efficient at producing fulllength cdna from long rna templates at temperatures between 4255. Contains protoscript ii reverse transcriptase, an enzyme with increased thermostability and reduced rnase h activity. 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.

Refer to nanodrop protocol for instructions on use. The first is the difficulty in reaching the cap site in the first strand synthesis with the reverse transcriptase rt, the first enzyme involved in the preparation of a cdna library. Applications first strand cdna synthesis for rt pcr and real time rtpcr, see protocol on back page. Random 9mers, also provided with this kit, are efficient primers for the detection of multiple short rtpcr targets.

Revertaid reverse transcriptase 10,000 u50,000 u ep0441ep0442 revertaid h minus reverse transcriptase 10,000 u50,000 u ep0451ep0452 cdna synthesis kits revertaid first strand cdna synthesis kit 20 rxns100 rxns k1621k1622 revertaid h minus first strand cdna synthesis kit 20 rxns100 rxns k1631k1632. It can be used with either sequencespecific primers, polydt15 primers, or. For reliable first strand cdna synthesis from all rna sources. Thermo scientific revertaid reverse transcriptase rt is a recombinant mmulv rt. Perform efficient synthesis of fulllength first strand cdna up to kb with this genetically modified mmulv rt for routine reverse transcription. A control reaction without reverse transcriptase is recommended to examine the dna contamination in the samples. First strand cdna synthesis kit direct pcr reagents and.

The first strand cdna synthesis kit is used for the synthesis of the first strand cdna as the starting reaction for rtpcr. Usb firststrand cdna synthesis kit for realtime pcr. Thermo scientific revertaid reverse transcriptase 200 u. Researchers can choose from among a wide variety of commercial cdna synthesis kits. If the cdna is to be used for secondstrand synthesis by the.

The maxima first strand cdna synthesis kit for rtqpcr is capable of reproducible cdna synthesis from a wide range of total rna amounts 1 pg to 5 g at elevated temperatures 42 to 65c. A robust experimental design delivers a universal kit that is suitable for first. Thermo scientific ep0441 view more versions of this product catalog no. Hyperscripttm first strand synthesis kit is ideally organized for synthe sizing reaction of first strand cdna from purified mrna or total rna. First strand cdna synthesis kit can be used to synthesize the first strand cdna by reverse transcription rt at higher temperatures than the wild type mmulv to reach higher cdna yields for difficult rna transcriptions. Experience with revertaid rt reverse transcription kit. Readytouse master mix for cdna synthesis from total rna. Ribolock rnase inhibitor, supplied with the kit, effectively protects rna templates from degradation. Maxima first strand cdna synthesis kit for rtqpcr, with dsdnase. Revertaid first strand cdna synthesis kit fermentas and the last, said to be the best but not explained to me why it is so smarter race cdna amplification kit clontech. This kit is can be used in conjunction with shineprobe qpcr kits or with any other qpcr kit suitable for the application. Hyperscript first strand synthesis kit 1 bohemia genetics.

Revertaid first strand cdna synthesis kit is a complete system for efficient synthesis of first strand cdna from rna templates. Thermo scientific revertaid h minus first strand cdna synthesis kit is a complete system for efficient synthesis of first strand cdna from mrna or total rna reverse transcriptase, which has a point mutation that completely eliminates rnase h activity. The enzyme maintains activity at 4250c and is suitable for synthesis of cdna up to kb. 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.

If random 9mers are used, the first strand synthesis reaction must be incubated at. The maxima first strand cdna synthesis kit contains oligodt 18 and random hexamer primers to prime synthesis of first strand cdna. The kit uses revertaid reverse transcriptase rt, which has lower rnase h activity compared to amv reverse transcriptase. Thermo scientific reverse transcription thermo fisher. The reverse transcriptase rt enzyme synthesizes the new cdna strand at a site determined by the type of primer used. Thermo scientific revertaid rt reverse transcription kit 500. Transcriptor first strand cdna synthesis kit roche life science. The revertaid first strand cdna synthesis kit was purchased from fermentas international inc.

Following first strand cdna synthesis, the sample can be used directly for in vitro amplification using pcr or for second strand synthesis using the gublerhoffman method. Revertaid h minus reverse transcriptase maintains activity over a. It differs from the mmulv rt by its structure and catalytic properties. The rna pcr kit was purchased from takara biotechnology co.

The amv first strand cdna synthesis kit combines amv enzyme mix and amv reaction mix. First strand cdna synthesis kitsreverse transcriptionrtpcr. Firststrand cdna synthesis kit ge healthcare, 27926101. Firststrand cdna synthesis for quantitative rtpcr origene. Using the revertaid first strand cdna synthesis kit quantify we need 0. Synthesis kit is a complete system for efficient synthesis of first strand cdna from mrna or total rna templates. Rnas were isolated using the trizol reagent invitrogen, and cdnas were synthesized using a revertaid first strand cdna synthesis kit fermentas. Quantification of the g6pdh gene with the transcriptor first strand cdna synthesis kit and the lightcycler 2. First strand cdna synthesis kit is designed to generate fulllength first strand cdna from mrna templates. Figure 2 shows that cdna prepared with the marligens first strand cdna synthesis system provides significantly enhanced sensitivity compared to cdna prepared with a competitors kit. Each entry pointing to the first page of the merged file. The kit includes revertaid h minus reverse transcriptase, which has a point mutation that completely eliminates rnase h activity. The kit can provide high efficient polya modification and reverse transcription and efficiently synthesize the firststrand cdna from 20 pg2. The mmlv reverse transcriptase 1ststrand cdna synthesis kit is optimized for.

Our pdf merger allows you to quickly combine multiple pdf files into one single pdf document, in just a few clicks. Add the following reagents into a sterile, nuclease free tube on ice in the indicated order. Accuscript high fidelity 1st strand cdna synthesis kit agilent. Add the following components in the indicated order. The master mix kit is used for first strand cdna synthesis from rna. First strand cdna synthesis after thawing, mix and briefly centrifuge the components of the kit. Only four to ten genetic differences between the strains circulating in the us and the original virus that was isolated from patients in. The enzyme is capable of first strand cdna synthesis up to kb. For robust twostep rtpcr and rtqpcr of rna up to 14 kb. Thermo scientific maxima h minus doublestranded cdna synthesis kit is a complete system for efficient synthesis of doublestranded cdna from total rna or mrna. Revertaid h minus reverse transcriptase, ribolock rnase inhibitor, 5x reaction buffer, dntp mix, oligodt18 primer, random hexamer primer, control rna, control primers and nucleasefree water. Thermo scientific revertaid reverse transcriptase 200 ul. Coumestrol induces mitochondrial biogenesis by activating. Amv reverse transcriptase provides first strand cdna synthesis reactions with a broader optimal reaction temperature from 37c to 50c.

Some are designed strictly for first strand cdna synthesis. Thermo scientific revertaid h minus first strand cdna synthesis kit perform efficient synthesis of first strand cdna from rna templates with this complete synthesis system. For robust twostep rtpcr and rt qpcr of rna up to 14 kb. Instruction manual protoscript ii first strand cdna. A fraction of the first strand cdna product was used to amplify sequences specific for three different messenger rnas using 1x longamp taq 2x master mix. The allinone firststrand cdna synthesis kit includes a reverse transcriptase and a specialized set of reagents designed to yield firststrand cdna that is optimal for gene cloning, cdna library creation and quantitative pcr amplification. Astragalus membranaceus inhibits inflammation via phospho. Thermo scientific maxima first strand cdna synthesis kit for. After completion of cdna synthesis, use 12 l of the first strand reaction out of the 20 l for pcr amplification. The goscript reverse transcription system is a convenient kit that includes a reverse transcriptase and an optimized set of reagents for efficient synthesis of firststrand cdna optimized in preparation for pcr amplification. Efficient cdna synthesis by allinone firststrand cdna synthesis kit. Highly efficient at producing fulllength cdna from as.

Thermo scientific revertaid h minus first strand cdna synthesis kit is a complete system for efficient synthesis of first strand cdna from rna templates. The transcriptor first strand cdna synthesis kit is designed to reverse transcribe rna mrna, total rna, viral rna, and in vitrotranscribed rna from a variety of sources for the following applications. Negative control reactions were carried out with 1x mmulv reaction mix. If the first strand cdna synthesis 33 dnarna hybrid itself is the starting material for second strand synthe sis this volume 35, omit step 3 and proceed immediately to step 4. Thermo scientific revertaid h minus reverse transcriptase. Mix thoroughly by pipetting up and down several times. Mint2 cdna synthesis kit utilizes the specific features of mint reverse transcriptase mint rt. Sensifast cdna synthesis kit, 50 reactions, first strand. Thermo scientific revertaid h minus first strand cdna. The enzyme possesses an rnadependent and dnadependent polymerase activity and a rnase h activity specific to rna in rnadna hybrids which is significantly lower than that of avian. The goscript reverse transcription system is a convenient kit that includes a reverse transcriptase and an optimized set of reagents for efficient synthesis of first strand cdna optimized in preparation for pcr amplification. The first strand cdna can then be used as a template for pcr. Applications first strand cdna synthesis for pcr or rtpcr gene expression data validation by using rtpcr benefits active at temperatures up to 55 c.

The thermo scientific revertaid rt kit is a complete system for efficient synthesis of first strand cdna from mrna or total rna templates. Not recommended or intended for diagnosis of disease in humans or animals. Use 1 g of isolated mrna to generate first strand cdna for secondstrand synthesis and subsequent cloning reactions. The amv enzyme mix is an optimized blend of amv reverse transcriptase and a murine rnase inhibitor. Using the first strand cdna synthesis kit, rna is reverse transcribed into single stranded cdna. Thermo scientific revertaid rt kit is a complete system for efficient synthesis of first strand cdna from mrna or total rna templates.

Rna isolationpurification and firststrand cdna synthesis. Thermo scientific molecular biology workflow solutions. Lightcycler faststart dna masterplus sybr green i, was then used with primers for g6pdh to quantify geneexpression levels. Make a cdna synthesis reaction mix by adding the iscript reverse transcription supermix to the dnasetreated rna template according to the guidelines in table 4. Revertaid h minus first strand cdna synthesis kit is a complete system for efficient synthesis of first strand cdna from rna templates. The energicscript cdna synthesis kit is intended for cdna synthesis for twostep quantitative reverse transcriptionpcr qrtpcr applications, where amplicons are usually around 200 bp in length. Once all the options are properly set, click run to start the execution. The revertaid h minus first strand cdna synthesis kit contains. This primer mixture ensures high sensitivity in low copy number transcript detection assays. Total rna isolated from human placenta was used as template rna in reverse transcription reactions using the allinonetm firststrand cdna synthesis kit together with the oligodt primer.

First strand cdna synthesis kit thermo scientific fermentas first strand cdna synthesis kit is a complete system for the efficient synthesis of first strand cdna from rna templates. The kit is suitable for synthesis of cdna up to kb. Firststrand cdna synthesis depending on the intended use of the firststrand cdna, synthesis may be performed using different volumes of bulk firststrand cdna mix, and different types and amounts of primer and rna. Different amounts of total rna from k562 cells were reverse transcribed using random hexamer primers and the reaction conditions outlined in the pack insert of the kit. The transcriptor high fidelity cdna synthesis kit is designed to reverse transcribe rna with an increased fidelity cat. First strand cdna synthesis starts from the 3end cds adapter containing an oligodt sequence that anneals to polya stretches of rna.

Revertaid h minus reverse transcriptase does not degrade rna in rnadna hybrids during synthesis of the first strand cdna, and therefore high yields of fulllength cdna from long templates are obtained. Thermo scientific revertaid first strand cdna synthesis kit is a complete system for efficient synthesis of first strand cdna from rna templates. Oligodt primer, random primer or a sequencespecific primer. Product information thermo scientific possesses rna. Store the remaining first strand cdna sample at 20 c until use. 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. If the synthesis of second strand cdna is contemplated by the classi cal method this volume 34, rna must be hydrolyzed step 3. The first strand cdna product generated is up to 10 kb. Mix rna sample and primer dt 23 vn in two sterile rnasefree microfuge tubes.

Protocol after thawing, mix and briefly centrifuge the components of the kit. If the first strand cdna synthesis 33 dnarna hybrid itself is the starting material for second strand synthe sis this volume 35. The synthesis reaction can be completed in 15 to 30 minutes. Truscript first strand cdna synthesis kit product insert. 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.