Use 1 g of isolated mrna to generate first strand cdna for secondstrand synthesis and subsequent cloning reactions. Transcriptor first strand cdna synthesis kit roche. Using the fermentas first strand cdna synthesis kit quantify we need 0. Accuscript high fidelity 1st strand cdna synthesis kit agilent. First strand cdna synthesis kit direct pcr reagents and. The amv enzyme mix is an optimized blend of amv reverse transcriptase and a murine rnase inhibitor. Invitrogen superscript doublestranded cdna synthesis kit. First strand cdna synthesized with this system can be directly used as a template in pcr or realtime pcr. 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. Synthesis kit is a complete system for efficient synthesis of first strand cdna from mrna or total rna templates.
A unique blend of random hexamer and anchored oligo dt primers in the sensifast cdna synthesis mix provides optimal sensitivity and accuracy of firststrand. The reverse transcriptase rt provided with the 1st strand cdna synthesis kit is stratagene. These reagents should remain stable for at least 1 year in their unopened containers. The sensitivity of the kit is verified by the detection of gapdh transcript in 20 pg total rna after 35 cycles. The kit uses revertaid reverse transcriptase rt, which has lower rnase h activity compared to amv reverse transcriptase. Using the transcriptor first strand cdna synthesis kit, total rna was reverse transcribed by random hexamer priming. Thermo scientific maxima h minus first strand cdna synthesis. The cdna reaction product can be used directly in the next step without being purified. Mint2 cdna synthesis kit utilizes the specific features of mint reverse transcriptase mint rt. These are available as convenient pdf files online at procedure for firststrand cdna synthesis. Since there is no need to perform a separate dnase digestion and repurify your rna samples, your time and costs are saved.
Firststrand cdna synthesis for quantitative rtpcr origene. The reverse transcriptase rt enzyme synthesizes the new cdna strand at a site determined by the type. Truscript first strand cdna synthesis kit product insert. Thermo scientific revertaid rt kit is a complete system for efficient synthesis of first strand cdna from mrna or total rna templates. The reverse transcriptase rt enzyme synthesizes the new cdna strand at a site determined by the type of primer used. Rochecdnatranscriptor first strand cdna synthesis kit. Firststrand cdna synthesis kit is designed to generate fulllength firststrand cdna from mrna templates. Instruction manual protoscript ii first strand cdna.
First strand cdna synthesis after thawing, mix and briefly centrifuge the components of the kit. First strand cdna synthesis ab kit bridges lab protocols. Different amounts of total rna from k562 cells were reverse. For robust twostep rtpcr and rtqpcr of rna up to 14 kb. First strand cdna synthesis starts from the 3end cds. For robust twostep rtpcr and rt qpcr of rna up to 14 kb. Mmulv enzyme mix combines mmulv reverse transcriptase and murine rnase inhibitor, while mmulv reaction mix contains dntps and an optimized buffer. The nebnext rna first strand synthesis module is designed for synthesis of cdna from rna fragments using protoscript ii reverse transcriptase and random priming. First strand cdna synthesis kit is designed to generate fulllength first strand cdna from mrna templates.
Usb firststrand cdna synthesis kit for realtime pcr. Quantification of the g6pdh gene with the transcriptor first strand cdna synthesis kit and the lightcycler 2. C4361 cdna synthesis reagents 200l first strand 5x buffer 20g oligodt 15 primer 20g random hexameric primers. Not recommended or intended for diagnosis of disease in humans or animals. Secondstrand synthesis this protocol is suitable for synthesizing secondstrand cdna from the 20. Protoscript ii first strand cdna synthesis kit features two optimized mixes, protoscript ii enzyme mix and protoscript ii reaction mix. It can be used with either sequencespecific primers, polydt15 primers, or. The goscript reverse transcription system is a convenient kit that includes a reverse transcriptase and an optimized set of. Mmulv enzyme mix combines mmulv reverse transcriptase and murine. This robust reverse transcription kit allows a selection of firststrand priming strategies, oligodt primers only, random primers only, or userdesigned genespecific primers.
Since results will be normalized in the end, standardizing rna quantities at this step is unnecessary. 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. 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. Maxima h minus first strand cdna synthesis kit with dsdnase, in addition to the above listed components, contains dsdnase and 10x dsdnase buffer. The kit also contains two optimized primers for reverse transcription and nucleasefree water. On ice, add the following reagents in the order shown to the firststrand reaction tube. The kit uses revertaid reverse transcriptase rt, which has lower.
Avoid repeated freezethaw cycles for control neo pa rna, rnase. Accuscript high fidelity 1st strand cdna synthesis kit. Norgens truscript first strand cdna synthesis kit components should be stored at 20oc. Using the first strand cdna synthesis kit, rna is reverse transcribed into single stranded cdna. Description legene express 1st strand cdna synthesis system is optimized to synthesize high yield cdna from total or poly a. The recombinant thermo scientific ribolock rnase inhibitor, supplied with the kit, effectively protects rna from degradation at temperatures up to 55 c.
Revertaid first strand cdna synthesis kit fermentas and the last, said to be the best but not explained to me why it is so. The length of cdna achieved is verified by the detection of a 5. The enzyme maintains activity at 4250c and is suitable for synthesis of cdna up to kb. Human total rna from 1 pg to 1 g was reverse transcribed using the iscript cdna synthesis kit or the iscript select cdna synthesis kit and the firststrand primers indicated above. The enzyme mix combines protoscript ii reverse transcriptase and. Figure 1 is a summary of the two procedures that may be. The first strand cdna kit combines genomic dna elimination of rna samples and subsequent first strand cdna synthesis in one easy workflow to ensure accuracy of realtime rtpcr results. Msds documents are not included with product shipments. Legene express 1st strand cdna synthesis system, size. The kit uses revertaid reverse transcriptase rt, a.
For firststrand cdna synthesis twotube rtpcr pdf 381kb english. Truscript first strand cdna synthesis kit for mrna product. Thermo scientific revertaid first strand cdna synthesis kit is a complete system for efficient synthesis of first strand cdna from rna templates. Enjoy the benefits of a tested protocol and reagents of consistent quality. The cdna synthesis procedure is illustrated in fig. Synthesis of 1st strand cdna following polyadenylation, the rna is used as template to synthesize 1st strand cdna. This robust reverse transcription kit allows a selection of firststrand priming strategies. Revertaid h minus first strand cdna synthesis kit sinapse. Ribolock rnase inhibitor, supplied with the kit, effectively protects rna templates from degradation. The first strand cdna can then be used as a template for pcr. For reliable first strand cdna synthesis from all rna sources.
Marligens firststrand cdna synthesis system for quantitative rtpcr has been designed for the highest efficiency conversion of rna to cdna and is. Efficient cdna synthesis by allinone firststrand cdna synthesis kit. 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. Add 10 ug rna in a final volume of 20 ul to each well. Revertaid first strand cdna synthesis kit thermo fisher scientific.
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. Universal riboclone cdna synthesis system c4360 each system contains sufficient reagents to process up to 40g mrna and to perform 10 adaptor ligation reactions. The cdna synthesis protocol is optimized for reverse transcription of mrna and mirna templates. Thermo scientific maxima h minus first strand cdna. The applicability of the transcriptor first strand cdna synthesis kit to partially. Rna isolationpurification and firststrand cdna synthesis. The kit can provide high efficient polya modification and reverse transcription and efficiently synthesize the firststrand cdna from 20 pg2.
The reverse transcription efficiency of the allinone first strand synthesis kit is assessed by examining the amplification results of different genes or gene regions using the oligodt synthesized cdna prepared from the allinone firststrand cdna kit. Add the following components in the indicated order. Maxima h minus first strand cdna synthesis kit contains maxima h minus enzyme mix, oligodt 18 and random hexamer primers, 5x rt buffer, dntp mix, and nucleasefree water. The amv first strand cdna synthesis kit combines amv enzyme mix and amv reaction mix.
Truscript first strand cdna synthesis kit for mrna product insert. 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. Transcriptor first strand cdna synthesis kit roche life science. Oligodt primer, random primer or a sequencespecific primer.
The primescript first strand cdna synthesis kit contains all of the reagents necessary for synthesis of firststrand cdna from total or. L depctreated water 91 5x secondstrand reaction buffer 30 10 mm dntp mix 3 e. Using the transcriptor first strand cdna synthesis kit, total rna was. Omniscript reverse transcription handbook en print bookmark share omniscript reverse transcriptase for firststrand cdna synthesis twotube rtpcr pdf 381kb english format file size. 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. First strand cdna synthesis kit igscript intact genomics. It uses cloned moloney murine leukemia virus mmlv reverse transcriptase to synthesize first strand cdna from an mrna population. The reactions described in the superscript doublestranded cdna synthesis kit protocol are designed to convert 2550g total rna, or 0.
Protoscript ii first strand cdna synthesis kit neb. Primers are not included in the preassembled first strand reaction. First strand cdna synthesis was carried out in the presence of 1x protoscript ii reaction mix and 1x protoscript ii enzyme mix at 42c using 250 ng of jurkat total rna. Amv reverse transcriptase provides first strand cdna synthesis reactions with a broader optimal reaction temperature from 37c to 50c. 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. Synthesis of firststrand cdna using reverse transcriptase. Precautions and disclaimers this kit is designed for research purposes only. Omniscript reverse transcription handbook en qiagen. The iscript select cdna synthesis kit is a sensitive, flexible, and easytouse kit for the generation of firststrand cdna.
First strand cdna synthesis protocols thaw system components and put on ice. Fermentas first strand cdna synthesis kit is a complete system for the efficient synthesis of first strand cdna from rna templates. It can be used with either sequencespecific primers, polydt15 primers, or random primers, pdn6. 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. The performance of protoscript ii first strand cdna synthesis kit is tested in an rt reaction using jurkat total rna with primer dt23vn. Goscript reverse transcription system quick protocol fb114pdf. The enzyme mix combines protoscript ii reverse transcriptase and murine rnase inhibitor, while the reaction mix contains dntps and an optimized buffer.
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