Revertaid first strand cdna synthesis kit is a complete system for efficient synthesis of first strand cdna from rna templates. For my project i have extracted rna from mouse gingival cells and below are some samples ive extracted for a trial run rt pcr. 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. The cdna synthesis system uses an optimized onetube procedure, based on the method of gubler and hoffmann. Researchers can choose from among a wide variety of commercial cdna synthesis kits. For multiple reactions, prepare a master mix of components common. Thermo scientific revertaid h minus first strand cdna. However, amplification of some pcr targets 1 kb may require the removal of rna complementary to the cdna. The kit uses revertaid reverse transcriptase rt, which has lower rnase h activity compared to amv reverse transcriptase. Revertaid first strand cdna synthesis kit thermo fisher scientific. First strand cdna synthesis kit 100 x 20 l rxns from. Rt enhancer is included to remove contaminating dna, eliminating the need for dnase i.
Highcapacity cdna reverse transcription kits product. It uses cloned moloney murine leukemia virus mmlv reverse transcriptase to synthesize first strand cdna from an mrna population. 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. Rna isolationpurification and firststrand cdna synthesis. Thermo scientific revertaid h minus first strand cdna synthesis kit is a complete system for efficient synthesis of first strand cdna from rna templates. Thermo scientific maxima h minus doublestranded cdna. Reverse transcriptases rts use an rna template and a short primer complementary to the 3 end of the rna to direct the synthesis of the first strand cdna. First strand cdna synthesis kit 100 x 20 l rxns k1612 from. Ncode sybr greener mirna qrtpcr kit 10 polyadenylation 20 cdna synthesis 100 qpcr reactions mirqer100 ncode mirna firststrand cdna synthesis kit 10 polyadenylation 20 cdna synthesis 50 polyadenylation 100 cdna synthesis mirc10 mirc50 rediplate 96 ribogreen rna quantitation kit 96well plate 8. The high capacity cdna reverse transcription kit delivers extremely highquality, singlestranded cdna from 0. 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. The revertaid first strand cdna synthesis kit contains revertaid reverse transcriptase, ribolock rnase inhibitor, 5x reaction buffer, dntp mix, oligodt18 primer, random.
Transcriptor first strand cdna synthesis kit roche life science. Smarter pcr cdna synthesis kit clontech 634925 or 634926 kapa hifi pcr kit kapa biosystems kk2101 or kk2102 additional 5 pcr primer iia any oligo synthesis vendor 1. This cdna anneals to one of the primers leading to firststrand synthesis. Thermo scientific maxima first strand cdna synthesis kit for. Tetro cdna synthesis kit is optimized for rt reactions over a wide range of total rna concentrations 10 pg5. The thermo scientific maxima first strand cdna synthesis kit for rtqpcr is a convenient system optimized for cdna synthesis in 2step quantitative rtpcr rtqpcr applications. Maxima h minus first strand cdna synthesis kit and. Some are designed strictly for firststrand cdna synthesis. C, which enables the production of cdna from the rna sample by reverse transcription. Component 375u kit 1,500u kit thermoscriptz rt 15 ul 25 l 100 l 5x cdna synthesis buffer 500 l 2. Superscript iv reverse transcriptase user guide 3 february 201526 superscript iv firststrand cdna synthesis reaction the example procedure below shows appropriate volumes for a single 20l reverse transcription reaction.
Ribolock rnase inhibitor, supplied with the kit, effectively protects rna templates from degradation. Superscript ii reverse transcriptase thermo fisher scientific. The verso system achieves robust and sensitive reverse transcription through the combination of a high affinity rt enzyme, a unique rna priming method, and an optimized buffering system. Set sample type setting to other sample and the constant to 33. Thermo scientific reverse transcription thermo fisher. Firststrand cdna synthesis using superscript ii rt, continued the cdna can now be used as a template for amplification in pcr. Thermo scientific revertaid first strand cdna synthesis kit is a complete system for efficient synthesis of first strand cdna from rna templates. For the rna precipitated, most concentrations are around 10ngul with. The thermo scientific reverse transcription portfolio offers robust and reliable. The synthesis of dna from an rna template, via reverse transcription, produces complementary dna cdna. However, this step can be omitted in some cases unpublished results. The kit uses revertaid reverse transcriptase rt, a. Superscript vilo cdna synthesis kit product information sheet pub.
The kit utilizes our smart switching mechanism at 5 end of rna template technology coupled with pcr. 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. Reverse transcriptases rts use an rna template and a primer complementary. This kit contains the highly efficient reverse transcriptase revertra ace and a rt. The kit components are fully optimized to generate maximum yields of full. Rtpcr reverse transcription pcr reverse transcriptases. Universal rt primers recognize the universal sequences present on both the 5 and 3 extended ends of the mature mirnas. 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. Choose the best cdna synthesis kit for your application, with.
The kit contains oligo dt 18 and random hexamer primers. Thermo scientific verso cdna synthesis kit provides robust transcription of rna to create a complete cdna pool. This is recommended when the cdna synthesis reaction uses less than 10 ng of rna input. Take appropriate laboratory safety measures and wear gloves when handling. Selection of thermo scientific reverse transcriptases includes both genetically modified. Monsterscript reverse transcriptase is a thermostable reverse transcriptase that completely lacks rnase h activity.
First strand cdna synthesis kit direct pcr reagents and. It can be used with either sequencespecific primers, polydt15 primers, or random primers, pdn6. The kit uses maxima reverse transcriptase rt, an advanced enzyme derived by in vitro evolution of mmulv rt. When stored under these conditions, the kit components are stable for 1 year. C, which enables the production of cdna from the rna sample. The kit is suitable for synthesis of cdna up to kb. Compare first strand cdna synthesis kit 100 x 20 l rxns k1612 from thermo fisher scientific on. This gives the same relative representation in cdna templates, regardless of gene abundance, making it excellent for use in qpcr studies. Highquality rna can be obtained by isolating total rna. Random random primers ensure that the first strand synthesis occurs efficiently with all species of rna molecules present, including mrna and rrna.
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. Store the hicdna synthesis kit at 20c in a constanttemperature freezer. An overview of technical considerations when using. 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. 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. Quanta qscript cdna synthesis kit quanta qscript cdna supermix qiagen quantitect reverse transcription kit stratagene affinityscript qpcr cdna synthesis kit.
It can be used with either sequencespecific primers, polydt15 primers, or. Reverse transcriptases rts use an rna template and a short primer complementary to the 3 end of 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. 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. Toyobo has various kits for cdna synthesis based on the genetically improved mmlv reverse transcriptase revertra ace. The applied biosystems highcapacity cdna reverse transcription kit uses the random primer scheme for initiating cdna synthesis. Superscript vilo cdna synthesis kit product information.
Tetro cdna synthesis kit contains our highly sensitive mmlv reverse transcriptase, oligo dt 18 and random hexamer primers and all the necessary components to. Thermostable protector rnase inhibitor is included in the kit to protect rna from degradation at high reaction temperatures. For cdna synthesis and genomic dna elimination in rna. High capacity cdna reverse transcription kits the high capacity cdna reverse transcription kits for 200 and reactions contain all the reagents needed for reverse transcription rt of total rna to singlestranded cdna using a reaction size of 20 l. Thermo scientific fermentas first strand cdna synthesis kit is a complete system for the efficient synthesis of first strand cdna from rna templates. Synthesis kit is a complete system for efficient synthesis of first strand cdna from mrna or total rna templates. Products cdna synthesis kits toyobo biotech support.
Rna isolationpurification and firststrand cdna synthesis entered by kevin janes janes lab protocols 122816 2 wet pellets can be warmed for a few seconds at 37oc to increase evaporation smaller or broken pellets will need to have the remaining liquid evaporated off at 42. Thermoscriptz rt can generate cdna transcripts from 100 bp to 12 kb at temperatures ranging from 50c to 65c. The first strand cdna synthesis kit is used for the synthesis of the first strand cdna as the starting reaction for rtpcr. Reverse transcriptases rts use an rna template and a primer complementary to the rna to direct the synthesis of the first strand cdna. 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. Efficiency kits varied in their ability to efficiently generate cdna from different rna inputs linearity of cdna synthesis kits. Ncode mirna amplification system thermo fisher scientific.
Safety information the hicdna synthesis kit is for laboratory use only, not for drug, household or other uses. Components of thermo scientific revertaid first strand cdna synthesis kit. Figure 1 is a summary of the two procedures that may be. Thermo scientific revertaid rt reverse transcription kit. The verso system achieves robust and sensitive reverse transcription through the. Reverse transcription rt is a widely used application in molecular biology research for rna and expression analysis. 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. The revertra ace qpcr rt kit is an efficient and convenient kit to synthesize high quality cdnas for realtime pcr. For your convenience, reagents are available as kits or standalone products, depending on your needs. 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. Thermo scientific revertaid first strand cdna synthesis kit.
For your convenience, reagents are available as kits or standalone. We used iscript reverse transcription supermix biorad for cdna synthesis. The monsterscript 1ststrand cdna synthesis kit has been discontinued. The thermo scientific maxima h minus doublestranded cdna synthesis kit is a complete system for efficient synthesis of doublestranded cdna from total rna or mrna. 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 enzymes lack of rnase h activity contributes to its ability to make longer cdnas and more complete fulllength libraries of firststrand cdna. Im completely new to rna extraction, cdna synthesis and rt pcr. The enzyme maintains activity at 4250c and is suitable for synthesis of cdna up to kb. This step is very important in order to perform pcr since dna polymerase can act only on dna. Incorporation of universal primer binding sites in a singlestep during firststrand cdna synthesis eliminates the need for tedious secondstrand synthesis and adaptor ligation. It is important to use highquality rna for your cdna prepar ation. Anchored oligodt primers and random hexamers provide flexible rna priming methods for cdna synthesis. Protocol for cdna synthesis and qrtpcr cdna synthesis. In rtpcr, reverse transcriptase and an rna sample are used in addition to the standard pcr reagents.
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