Efficiency kits varied in their ability to efficiently generate cdna from different rna inputs linearity of cdna synthesis kits with bactin 15 20 25 30 35 401. The thermo scientific revertaid rt kit is a complete system for efficient synthesis of first strand cdna from mrna or total rna templates. Unesco eolss sample chapters genetics and molecular biology reverse transcriptase and cdna synthesis kunitada shimotohno encyclopedia of life support systems eolss 3. The resulting singlestranded cdna is suitable for use in realtime quantitative reverse transcription pcr rtqpcr. The kit provides all reagents required for firststrand cdna synthesis reactions. This gives the same relative representation in cdna templates, regardless of gene abundance, making it excellent for use in qpcr studies. The revertaid first strand cdna synthesis kit contains revertaid reverse transcriptase, ribolock rnase inhibitor, 5x reaction buffer, dntp mix, oligodt18 primer, random hexamer primer, control gapdh rna, 10. The latter is designed to bind at the beginning of the polya tail to generate fulllength. First strand cdna synthesis kit, supplied by thermo fisher, used in various techniques. The protocol described here used the isolate rna mini kit columns systems.
Nzy firststrand cdna synthesis kit is a system that includes all the necessary components to synthesize firststrand cdna, except the template rna. This robust reverse transcription kit allows a selection of firststrand priming strategies, oligodt primers only, random primers only, or userdesigned genespecific primers. Rtpcr kits, assays and reagents reverse transcriptases. Nzy firststrand cdna synthesis kit is formulated to provide high yields of fulllength cdna products and to increase. Nucleasefree water can be stored at room temperature. It is important to use highquality rna for your cdna prepar ation. Thermo scientific maxima first strand cdna synthesis kit. Thermo scientific reverse transcription thermo fisher. Tetro cdna synthesis kit contains our highly sensitive mmlv reverse transcriptase, oligo dt 18 and random hexamer primers and all the necessary components to generate high quality cdna from rna templates fig. A balanced amount of oligo dtand random primers is.
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. 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. Thermo scientific maxima first strand cdna synthesis kit for. January 2017 for the synthesis of doublestranded cdna from total rna or mrna cat. Includes goscript reverse transcriptase, mmlv and amv reverse transcriptases. 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. Nzy firststrand cdna synthesis kit cdna kits nzytech. Rna was reverse transcribed using a firststrand cdna synthesis kit fermentas. Rna isolationpurification and firststrand cdna synthesis. The sensifast cdna synthesis kit provides a rapid and sensitive method for firststrand cdna synthesis, which displays excellent linearity across a wide range of starting material.
Optimizing a qpcr gene expression quantification assay for s. The iscript select cdna synthesis kit is a sensitive, flexible, and easytouse kit for the generation of firststrand cdna. 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. Strand cdna synthesis kit with random hexamer primers fermentas, lithuania. The tonkbio first chain cdna synthesis kit for rtpcr is optimized to efficiently synthesize first chain cdna from mrna or total rna templates. The kit is suitable for synthesis of cdna up to kb. The recombinant thermo scientific ribolock rnase inhibitor, supplied with the kit, effectively protects rna from degradation at temperatures up to 55c. Maxima sybr green master mix fermentas, iq sybr green. 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.
Omniscript reverse transcriptase for firststrand cdna synthesis twotube rtpcr pdf 381kb english format file size language download get adobe reader contact qiagen. The revertaid first strand cdna synthesis kit is supplied with both oligodt 18. We recommend incubation at 42c for one hour for maximum yield and length. Importance of the efficiency of doublestranded dna formation. Ribolock rnase inhibitor, supplied with the kit, effectively protects rna templates from degradation. This technology is also becoming important in clinical diagnostics, such as the monitoring of gene expression profiles in breast cancer 16. Thermo scientific maxima h minus doublestranded cdna synthesis kit for 10 reactions dna synthesis reagents and columns biochemicals and reagents. Reverse transcription rt reaction is a process in which singlestranded rna is reverse transcribed into complementary dna cdna by using total cellular rna or polya rna, a reverse transcriptase enzyme, a primer, dntps and an rnase inhibitor. The verso enzyme cdna synthesis kit is widely used for cdna synthesis, due to its proven ability to produce high yields of fully representative full length cdna from low yields of rna. First strand cdna synthesis kit direct pcr reagents and kits. Sugarcane transcript profiling assessed by cdnaaflp. Quanta qscript cdna synthesis kit quanta qscript cdna supermix qiagen quantitect reverse transcription kit stratagene affinityscript qpcr cdna synthesis kit. The firststrand cdna generated is ideal for pcr fig.
Enzymes for cdna synthesis compare and order genomics. I use fermentas themoscientific first strand cdna synthesis kit k1622 for cdna synthesis and use both of them for cdna synthesis that included. Superscript iii firststrand synthesis system for rtpcr. First strand cdna synthesis kit thermo fisher bioz ratings for. The thermo scientific reverse transcription portfolio offers robust and reliable reverse transcriptases in a variety of optimized formats for efficiency in rtpcr and rtqpcr applications. Add the following components in the indicated order.
The choice of the adapter pair to use for a particular cdna preparation depends on intended application. Mint2 cdna synthesis kit contains different plugoligo and cds adapters allowing synthesis of cdna with different flanking sequences. Protocol for cdna synthesis and qrtpcr cdna synthesis. First strand cdna synthesis kit 100 x 20 l rxns from. Kits whose sixdigit lot number begins with a 2 are not. Set sample type setting to other sample and the constant to 33. The kit uses mmlv reverse transcriptase with lower rnase h activity, which avoids degradation of rna when rnadna hybrid forms in. The kit is formatted for highyield production of cdna up to 12 kb and each reaction accommodates a range of rna amounts from fg to g. 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. Hi, in our lab we study gene expression in plants and good rna and cdna synthesis is very important. Supplied with the recombinant ribolock rnase inhibitor. Using the fermentas first strand cdna synthesis kit quantify we need 0.
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 uses revertaid reverse transcriptase rt, which has lower rnase h activity compared to amv reverse transcriptase. Impairment of hiv1 cdna synthesis by dbr1 knockdown. First strand cdna synthesis kit 100 x 20 l rxns k1612 from.
Guaranteed for 12 months at 20c in a constant temperature freezer. Goscript uses mmlv reverse transcriptase and stateoftheart buffer to drive robust, reliable cdna synthesis of a full range of rare and abundant transcripts. Omniscript reverse transcriptase for firststrand cdna synthesis twotube rtpcr pdf 381kb english format. First strand cdna synthesized with this system can be directly used as a template in pcr or realtime pcr. For your convenience, reagents are available as kits or standalone products, depending on your needs. Despite new technologies for sample preparation, array treatment, and data analysis, concerns have been raised regarding the quality of the experimental.
Reverse transcription rt is a widely used application in molecular biology research for rna and expression analysis. Rt reaction is also called first strand cdna synthesis. Thermo scientific revertaid first strand cdna synthesis kit is a complete system for efficient synthesis of first strand cdna from rna templates. Activation of defense against phytophthora infestans in potato by downregulation of syntaxin gene expression lennart eschenlippold1, ramona landgraf1, ulrike smolka1, sebastian schulze2, mareike heilmann3, ingo heilmann3, gerd hause4 and sabine rosahl1 1department of stress and developmental biology, leibniz institute of plant biochemistry, weinberg 3, d06120 halle saale, germany. Two cdna synthesis primers are provided with the kit. Thermo scientific revertaid rt reverse transcription kit 500 x. The first and the second strands cdna synthesis was performed with the revertaidtm h minus first strand cdna synthesis kit fermentas, lithuania following the manufacturers instructions. The kit components are fully optimized to generate maximum yields of full. The synthesis reaction can be completed in 15 to 30 minutes. For priming, three different primer systems can be used. Add 498 ul of up ultra pure water into a microcentrifuge tube 2. 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. Revertaid first strand cdna synthesis kit thermo fisher scientific.
Thermo scientific revertaid first strand cdna synthesis kit. Other factors required for reverse transcription metal ions such as magnesium and manganese ions are essential for the reaction, and. Monarch nucleic acid purification kits are optimized for maximum performance and minimal environmental impact. Apr 01, 20 the concentration of cdna after the secondstrand dna synthesis was measured with a nanodrop nd spectrophotometer nanodrop technologies. 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. Selection of the proper cdna synthesis kit is critical for detailed analysis of quantitative gene expression. The thermo scientific reverse transcription portfolio offers robust and reliable reverse transcriptases in a variety of optimized formats for. Verso cdna synthesis kit from thermo scientific biocompare.
Increase the temperature of firststrand reaction up to 55c. Thermo scientific maxima h minus doublestranded cdna. Tetro cdna synthesis kit is optimized for rt reactions over a wide range of total rna concentrations 10 pg5. Highquality rna can be obtained by isolating total rna. A complete system for efficient synthesis of first strand cdna from rna templates. Importance of the efficiency of doublestranded dna. Thermo scientific revertaid rt reverse transcription kit. Compare first strand cdna synthesis kit 100 x 20 l rxns k1612 from thermo fisher scientific on. Omniscript reverse transcription handbook en qiagen. Rnaquant cdna reaction setup for 1 rna sample to be assayed on qpcr 96well plates it is important to start with total rna that includes the lncrna fraction. Please refer to the section below to choose the pair of adapters suitable for your needs.
However, this step can be omitted in some cases unpublished results. The kit is formatted for highyield production of cdna up to 12 kb and each reaction accommodates a. 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. The enzyme maintains activity at 4250 c and is suitable for. Analysis of gene expression with microarrays is a major application in functional genomics. Rtpcr kits and standalone reverse transcriptase enzymes for reverse transcription of fulllength cdna from your experimental sample. May 15, 20 selection of the proper cdna synthesis kit is critical for detailed analysis of quantitative gene expression. 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. The kit contains oligo dt 18 and random hexamer primers. Efficient synthesis of first strand cdna up to 9 kb.
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