Marligens firststrand cdna synthesis system for quantitative rtpcr has been designed for the highest efficiency conversion of rna to cdna and is fully optimized for quantitative realtime pcr applications. Protoscript ii first strand cdna synthesis kit features two optimized mixes, protoscript ii enzyme mix and protoscript ii reaction mix. First strand cdna synthesis kit igscript intact genomics. For robust twostep rtpcr and rtqpcr of rna up to 14 kb. Firststrand cdna synthesis kit ge healthcare, 27926101. Smarter cdna synthesis all commonly used cdna synthesis methods rely on the ability of reverse transcriptase rt to transcribe mrna into singlestranded ss dna in the first strand reaction. Find product specific information including cas, msds, protocols and references.
The applicability of the transcriptor first strand cdna synthesis kit to partially degraded ffpet rna was evaluated and compared with the widely used first strand cdna synthesis kit for rtpcr amv. Protoscript ii reverse transcriptase is a recombinant mmulv reverse transcriptase with reduced rnase h. The thermo scientific reverse transcription portfolio offers robust and reliable. 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 performance of protoscript first strand cdna synthesis kit is tested in an rt reaction using human jurkat total rna with primer dt 23vn.
Firststrand cdna synthesis for quantitative rtpcr origene. The length of cdna achieved is verified by detection of a 9. 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. The transcriptor high fidelity cdna synthesis kit is designed to reverse transcribe rna with an increased fidelity compared to other reverse transcriptases.
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. Efficient synthesis of fulllength first strand cdna up to kb optimum activity at 42c active up to 50c incorporates modified nucleotides e. First strand cdna synthesis kit direct pcr reagents and. Researchers can choose from among a wide variety of commercial cdna synthesis kits. 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. First strand cdna synthesis kit for rtpcr amv sigmaaldrich. The generated cdna is suitable for pcr with genespecific primers or for other downstream applications. 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.
Performance of the transcriptor first strand cdna synthesis kit in twostep rtpcr. Asfs002 product summary product description the rtstar first strand cdna synthesis kit 3 adaptor is designed to create cdna. Protocol for cdna synthesis and qrtpcr cdna synthesis. Synthesis kit is a complete system for efficient synthesis of first strand cdna from mrna or total rna templates. Thermo scientific revertaid reverse transcriptase 200 ul.
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. Secondstrand synthesis this protocol is suitable for synthesizing secondstrand cdna from the 20. The first strand cdna product generated is up to 10 kb. Revertaid first strand cdna synthesis kit 100 x 20 l rxns. Reverse transcription of rna into cdna protocol materials. Smart switching mechanism at 5 end of rna template is a unique technology that allows the efficient incorporation of known sequences at both ends of cdna during first strand synthesis, without adapter ligation. Betaine reduces pausing and stops by the reverse transcriptase, enabling improved reverse transcription through difficult sequences, such as regions of high gc content. A control reaction without reverse transcriptase is recommended to examine the dna contamination in. Thermo scientific revertaid rt 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. Invitrogen cloned amv firststrand cdna synthesis kit. Thermo scientific revertaid first strand cdna synthesis kit 100 reactions life sciences. Components of thermo scientific revertaid first strand cdna synthesis kit.
Have you tried the first strand cdna synthesis kit by promega. For use with application firststrand cdna synthesis, rtpcr. Shop online for a wide selection of thermo scientific revertaid first strand cdna synthesis kit perform efficient synthesis of first strand cdna from rna. The kit is suitable for synthesis of cdna up to kb. The kit can provide high efficient polya modification and reverse transcription and efficiently synthesize the firststrand cdna from 20 pg2. Maxima first strand cdna synthesis kit for rtqpcr, with. The first strand of cdna can be directly used as a template in pcr see rtpcr protocol or in second strand cdna synthesis see protocol. The kit uses revertaid reverse transcriptase rt, which. Using the revertaid first strand cdna synthesis kit. Revertaid first strand cdna synthesis kit fermentas and the last, said to be the best but not explained to me why it is so. First strand cdna synthesis kit is designed to generate fulllength first strand cdna from mrna templates. The iscript select cdna synthesis kit is a sensitive, flexible, and easytouse kit for the generation of firststrand cdna. The tonkbio first chain cdna synthesis kit for rtpcr is optimized to efficiently synthesize first chain cdna from mrna or total rna templates.
Sigmaaldrich offers roche11483188001, first strand cdna synthesis kit for rtreverse transcriptasepcr amv for your research needs. Revertaid first strand cdna synthesis kit sigmaaldrich. This robust reverse transcription kit allows a selection of firststrand priming strategies, oligodt primers only, random primers only, or userdesigned genespecific primers. The enzyme mix combines protoscript ii reverse transcriptase and murine rnase inhibitor, while the reaction mix contains dntps and an optimized buffer.
Therefore, degradation of rna does not occur during first strand. Mmulv enzyme mix combines mmulv reverse transcriptase and murine rnase inhibitor, while mmulv reaction mix contains dntps and an optimized buffer. The first strand of cdna can be directly used as a template in pcr see rtpcr protocol, realtime pcr or in second strand cdna synthesis see protocol. Thermo scientific molecular biology workflow solutions. Truscript first strand cdna synthesis kit product insert. The enzyme mix combines protoscript ii reverse transcriptase and.
Thermo scientific fermentas first strand cdna synthesis kit is a complete system for the efficient synthesis of first strand cdna from rna templates. Transcriptor first strand cdna synthesis kit roche. Ribolock rnase inhibitor, supplied with the kit, effectively protects rna templates from degradation. Thermo scientific revertaid reverse transcriptase perform efficient synthesis of fulllength first strand cdna up to kb with this genetically modified mmulv rt for routine reverse transcription. L depctreated water 91 5x secondstrand reaction buffer 30 10 mm dntp mix 3 e. Thermo scientific revertaid rt reverse transcription kit. Transcriptor first strand cdna synthesis kit roche life science. Primers are not included in the preassembled first strand. On ice, add the following reagents in the order shown to the firststrand reaction tube.
Some are designed strictly for first strand cdna synthesis. The kit uses revertaid reverse transcriptase rt, which has lower rnase h activity compared to amv reverse transcriptase. Random hexamer primer 5 x reaction buffer ribolock rnase inhibitor 20 ul 10 mmdntp mix revertaid mmilv rt 200 ul water, nuclease free 0. Revertaid first strand cdna synthesis kit thermo fisher scientific. Thermo scientific reverse transcription thermo fisher.
Thermo scientific revertaid first strand cdna synthesis kit 100. It provides a dramatically simplified workflow that combines genomic dna elimination and cdna synthesis into one. Thermo scientific revertaid first strand cdna synthesis kit is a complete system for efficient synthesis of first strand cdna from rna templates. Primers are not included in the preassembled first strand reaction. Total rna was reverse transcribed using the anchoredoligodt 18 primer and reaction conditions outlined in the pack insert of the product. The enzyme maintains activity at 4250 c and is suitable for.
In the subsequent pcr, 5 l of the cdna reaction was used. Accuscript high fidelity 1st strand cdna synthesis kit. Protoscript first strand cdna synthesis kit features two optimized mixes, mmulv enzyme mix and mmulv reaction mix. The kit uses revertaid reverse transcriptase rt, a recombinant mmulv rt which maintains activity at 4250c and is suitable for synthesis of cdna. Add the following components in the indicated order.
The thermo scientific revertaid rt kit is a complete system for efficient synthesis of first strand cdna from mrna or total rna templates. However, because rt cannot always transcribe the entire mrna sequence, the 5 ends of genes tend to be underrepresented in cdna. Reverse transcription rt is a widely used application in molecular biology research for rna and expression analysis. Revertaid first strand cdna synthesis kit is a complete system for efficient synthesis of first strand cdna from rna templates. The sensitivity of the kit is verified by the detection of gapdh transcript in 20 pg total rna after 35 cycles. 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. 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 kit uses revertaid reverse transcriptase rt, which has lower. 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.
Using the revertaid first strand cdna synthesis kit quantify we need 0. 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. Thermo scientific revertaid premium first strand cdna. The presence of these known sequences is crucial for a number of downstream applications, including amplification, race, and library. Compare revertaid first strand cdna synthesis kit 100 x 20 l rxns k1622 from thermo fisher scientific on. This kit is powered by primescript reverse transcriptase primescript rt, which has exceptionally strong strand displacement activity and efficiently synthesizes cdna. First strand cdna synthesis protocols e6300 introduction. Thermo scientific revertaid first strand cdna synthesis kit. 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. Search results for revertaid first strand cdna synthesis kit at sigmaaldrich. Mar 07, 2011 cdna synthesis exploits the viral enzyme reverse transcriptase rt, which uses mrna as a template to drive dna synthesis, a step called reverse transcription.