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