Using the maxima first strand cdna synthesis kit for rtqpcr as the reference, the. The following is intended as a guideline an d starting point when using first strand cdna in pcr with taq dna polymerase. Reverse transcription rt is a widely used application in molecular biology research for rna and expression analysis. We recommend incubation at 42c for one hour for maximum yield and length. Includes maxima h minus reverse transcriptase is supplied with.
Expression analysis of st3gal4 transcripts in cervical cancer cells. Superscript iv reverse transcriptase is available as standalone enzyme or in a firststrand synthesis system format. Ribolock rnase inhibitor, supplied with the kit, effectively protects rna templates from degradation. Thermo scientific fermentas first strand cdna synthesis kit is a complete system for the efficient synthesis of first strand cdna from rna templates. Protocol for cdna synthesis and qrtpcr cdna synthesis. First strand cdna synthesis kit for rtpcr amv sigmaaldrich. N8080127 protoscript ii first strand cdna synthesis kit neb, cat. This kit is powered by primescript reverse transcriptase primescript rt, which has exceptionally strong strand displacement activity. 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.
Compare first strand cdna synthesis kit 100 x 20 l rxns k1612 from thermo fisher scientific on. 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. Firststrand cdna synthesis kit is designed to generate fulllength firststrand cdna from mrna templates. 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.
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. Maxima h minus cdna synthesis master mix was compared to thermo scientific maxima first strand cdna synthesis kit for rtqpcr and master mixes from other suppliers using 96gene applied biosystems taqman assay panels with 100 ng hela total rna input. 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. Thermo scientific revertaid first strand cdna synthesis. 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. Thermo scientific revertaid h minus first strand cdna synthesis kit is a complete system for efficient synthesis of first strand cdna from rna templates. Biorads iscript reverse transcription rt kits provide fast and sensitive firststrand cdna synthesis. The first strand cdna product generated is up to 10 kb. After synthesis, the cdna can be amplified with specific primers by pcr without int. Mirna cdna synthesis kit, with polya polymerase tailing. Maxima first strand cdna synthesis kit for rtqpcr, with. The thermo scientific reverse transcription portfolio offers robust and reliable. Quantification of the g6pdh gene with the transcriptor first strand cdna synthesis kit and the lightcycler 2. 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.
Thermo scientific maxima h minus first strand cdna. Thermo scientific revertaid rt reverse transcription kit. Thermo scientific revertaid first strand cdna synthesis kit, 100 reactions, price. The kit uses mmulv vity compared to amw reverse transcriptase. Transcriptor first strand cdna synthesis kit roche life science. Protoscript ii first strand cdna synthesis kit neb. Sybr is a trademark of thermo fisher scientific inc. Thermo scientific revertaid rt kit is a complete system for efficient synthesis of first strand cdna from mrna or total rna templates. Transcriptor reverse transcriptase is recommended for rtpcr because of its high sensitivity in combination with high thermostability. The kit uses the maxima h minus reverse transcriptase rt, which is an. It can be used with either sequencespecific primers, polydt 15 primers, or random primers. Thermo scientific maxima h minus reverse transcriptase 200.
Revertaid first strand cdna synthesis kit is a complete system for efficient synthesis of first strand cdna from rna templates. Components of thermo scientific revertaid first strand cdna synthesis kit. Reverse transcriptase, which has lower rnase h acti first. First strand cdna synthesis kit is designed to generate fulllength first strand cdna from mrna templates. Sensifast cdna synthesis kit, 50 reactions, first strand. Using the fermentas first strand cdna synthesis kit quantify we need 0. Thermo scientific revertaid first strand cdna synthesis kit is a complete system for efficient synthesis of first strand cdna from rna templates. The firststrand synthesis system includes dntps, oligodt20, random hexamers, control primers, and required buffer components. Get highly efficient synthesis of first strand cdna with the thermo scientific maxima h minus first strand cdna synthesis kit. Increase the temperature of firststrand reaction up to 55c.
Thermo scientific maxima h minus first strand cdna synthesis. Thermo scientific maxima h minus reverse transcriptase. Large fragments 7kb with thermo scientific maxima h minus. It provides a dramatically simplified workflow that combines genomic dna elimination and cdna synthesis into onetube procedure. The enzyme maintains activity at 37c and is suitable for synthesis of cdna up to 9 kb. Anchored oligodt primers and random hexamers provide flexible rna priming methods for cdna synthesis. Reverse transcriptases rts use an rna template and a primer complementary. Rt enhancer is included to remove contaminating dna, eliminating the need for dnase i. However, this step can be omitted in some cases unpublished results. The system can be used with as little as 1 ng or as much as 5 g of total rna.
Therefore, degradation of rna does not occur during first strand. First strand cdna synthesis kit 100 x 20 l rxns k1612 from. Random hexamer primer 5 x reaction buffer ribolock rnase inhibitor 20 ul 10 mmdntp mix revertaid mmilv rt 200 ul water, nuclease free 0. The kit uses revertaid reverse transcriptase rt, a. Page 2 of 4 firststrand cdna synthesis using thermoscriptz rt 1. 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. Add the following components to a nucleasefree microcentrifuge tube. 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 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. Firststrand cdna synthesis kit ge healthcare, 27926101. The kit includes superscript iv reverse transcriptase rt, a proprietary mmlv mutant that produces robust and reliable cdna synthesis in rt reactions.
Superscript iii firststrand synthesis system for rtpcr. Thermo scientific reverse transcription thermo fisher. All necessary components are in separate tubes for maximum optimization of cdna synthesis and to. The kit uses revertaid reverse transcriptase rt, which has lower rnase h activity compared to amv reverse transcriptase. Sensifast cdna synthesis kit, 50 reactions, first strand synthesis, fulllength reverse transcription. The firststrand synthesis system includes dntps, oligodt20, random hexamers. 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. Thermo scientific revertaid rt reverse transcription kit 500.
Maxima first strand cdna synthesis kit for rtqpcr with dsdnase. Purity and quality of rna was determined by spectrophotometer thermo scientific, san jose, california, usa and 1% agarose gel electrophoresis. The revertaid first strand cdna synthesis kit contains revertaid reverse transcriptase, ribolock rnase inhibitor, 5x reaction buffer, dntp mix, oligodt18 primer, random. Revertaid first strand cdna synthesis kit thermo fisher scientific. Thermoscript reverse transcriptase 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. Truscript first strand cdna synthesis kit product insert. Superscript iv reverse transcriptase from thermo fisher. The kit uses the maxima h minus reverse transcriptase rt, which is an advanced enzyme derived by in vitro evolution of mmulv rt. Thermo scientific dynamo flash probe qpcr kit f455s cdna synthesis thermo scientific maxima first strand cdna synthesis kit for rtqpcr ferk1641 thermo scientific maxima first strand cdna.
Depending on the length of template and degree of secondary structure, the efficiency of the first strand synthesis maybe improved by optimizing temperature and time 42. Following firststrand cdna synthesis, the sample can be used directly for in vitro. 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 kit includes revertaid h minus reverse transcriptase, which has a point mutation that comple. Super script vilo invitrogen, revertaaid first strand cdna synthesis. Thermo scientific first strand cdna synthesis kit is a complete system for efficient synthesis of first strand cdna from mrna or total rna templates.
Random hexamer primer 5 x reaction buffer ribolock rnase. Firststrand cdna synthesis was performed with a revertaid h minus first strand cdna synthesis kit thermo fisher scientific, inc. Scientific revertaid first strand cdna synthesis kit. William smith general phone 2755731 molecular biology sales specialist. First strand cdna synthesis is tested in twostep rt. The first strand cdna synthesis kit is used for the synthesis of the first strand cdna as the starting reaction for twostep rt pcr. U of rochester biological supply ctr g8633 stock manager. Thermo scientific revertaid h minus first strand cdna. The kit is suitable for synthesis of cdna up to kb.
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