Transcriptor first strand cdna synthesis kit roche pdf

This robust reverse transcription kit allows a selection of firststrand priming strategies, oligodt primers only, random primers only, or userdesigned genespecific primers. Highly efficient multiplex human t cell engineering. First strand cdna synthesis for qrtpcr realtime pcr. To prepare total rna or mrna, we recommend using roche life science reagents. Herein, we evaluated six commercially available kits for the synthesis of cdna according to amplification success rate, linearity and abl1 copy number.

No matter which pcr instrument you are using, this master mix, supplied in just two vials, brings out the full potential of your qrt. Oligo dt1218 nucleotides are most frequently used to initiate first strand cdna synthesis, if fulllength cdna synthesis is required. Always run at least one notemplate control ntc replace the template dna with water. I come across two kit which is the transcriptor first strand cdna synthesis kit and 1st strand cdna synthesis kit for rtpcr amv. Monsterscript reverse transcriptase is a thermostable reverse transcriptase that completely lacks rnase h activity. Cellulysertm lysis and cdna synthesis kit version 1. Which kit for cdna synthesis do you recommend to be used. Make sure that the genespecific primer is able to bind to the mrna antisense direction. Neb offers several reagents for cdna synthesis for use in applications including qpcr and qrtpcr. The cdna reaction product can be used directly in the next step without being purified.

Transcriptor universal cdna master fast cdna synthesis for. 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. Synthesis of first strand cdna for use in subsequent amplification reactions rtpcr on thermal block cyclers or. Isolated rna was subjected to cdna synthesis using the transcriptor first strand cdna synthesis kit. The transcritpor first strand cdna synthesis kit by roche has been specifically designed to reverse transcribe a variety of rna species mrna, total rna, viral rna and in vitro transcribed rna from a variety of. Transcriptor first strand cdna synthesis kit roche. Transcriptor high fidelity cdna synthesis kit version november 2008 cat. This is the first time that i will be peforming qrtpcr. For a selection of products which produce high quality intact rna templates. Roche diagnostics transcriptor first strand cdna synthesis kit, 100 reactions. 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.

Roche diagnostics has made every reasonable effort to ensure that all the information contained in this manual. Roche diagnostics transcriptor first strand cdna synthesis kit, 100. Roche diagnostics high pure rna isolation kit combined with roche diagnostics transcriptor first strand cdna synthesis kit. The performance of three cdna synthesis kits was com. Accurate cdna levels for unbiased twostep qrtpcr article pdf, 196 kb.

The enzymes lack of rnase h activity contributes to its ability to make longer cdnas and more complete fulllength libraries of firststrand cdna. Rna was reverse transcribed to cdna using the transcriptor first strand cdna synthesis kit with both random hexamers and oligo dt primers roche, basel, switzerland. 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. For priming, three different primer systems can be used. Transcriptor 1st strand cdna synthesis kit 04 379 012 001 for 50.

For robust twostep rtpcr and rtqpcr of rna up to 14 kb. Transcriptor high fidelity cdna synthesis kit sigmaaldrich. Gene expression analysis in biomarker research and early drug. One microgram of the total rna was reverse transcribed using a transcriptor firststrand cdna synthesis kit roche applied science, penzberg, germany, and. Transcriptor reverse transcriptase is a fast, recombinant reverse transcriptase expressed in e. For robust twostep rtpcr and rt qpcr of rna up to 14 kb. Which kit for cdna synthesis do you recommend to be used on bacterial rna from soils. Transcriptor first strand cdna synthesis kit from roche. Roche diagnostics transcriptor first strand cdna synthesis.

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. This kit is powered by primescript reverse transcriptase primescript rt, which has exceptionally strong stranddisplacement activity and efficiently synthesizes cdna. Fermentas first strand cdna synthesis kit is a complete system for the efficient synthesis of first strand cdna from rna templates. First strand cdna synthesis kit for rtpcr and the transcriptor first strand cdna synthesis kit achieved 15 successful reads at the 0.

First strand cdna synthesis protocols e6300 introduction. Mmulv enzyme mix combines mmulv reverse transcriptase and murine rnase inhibitor, while mmulv reaction mix contains dntps and an optimized buffer. The kit can provide high efficient polya modification and reverse transcription and efficiently synthesize the firststrand cdna from 20 pg2. The transcriptor first strand cdna synthesis kit includes transcriptor reverse transcriptase, an enzyme which achieves a broad dynamic detection range from very low to very high copy numbers of in vitro transcribed rna 3. Rna isolationpurification and firststrand cdna synthesis. Study gene expression levels, via twostep rtpcr, using qualitative rtpcr on conventional thermal cyclers or. Roche diagnostics high pure viral nucleic acid kit combined with roche diagnostics transcriptor first strand cdna synthesis kit. Transcriptor first strand cdna synthesis kit roche life science. Maxima h minus first strand cdna synthesis kit with dsdnase contains maxima h minus enzyme mix, dsdnase, 10x dsdnase buffer, oligodt 18 and random hexamer primers, 5x rt buffer, dntp mix, and nucleasefree water. Truscript first strand cdna synthesis kit product insert. Transcriptor reverse transcriptase the core component of the kit is a.

The latter is designed to bind at the beginning of. Applicability of function tested realtime ready assays for gene expression analysis article pdf, 1. The following conditions describe a firststrand cdna synthesis for a. Maxima h minus first strand cdna synthesis kit, with dsdnase. A control reaction without reverse transcriptase is recommended to examine the dna contamination in the samples. Mix rna sample and primer dt 23 vn in two sterile rnasefree microfuge tubes. Rtpcr is also used for the creation of cdna libraries, molecular cloning by pcr, and sequencing. High pure ffpe rna micro kit or the magnapure lc rna isolation kit iii tissue on the mplc 2. Briefly centrifuge them before starting the procedure. Reverse transcription is performed using the transcriptor first strand cdna synthesis kit roche. For your convenience, reagents are available as kits or standalone products, depending on your needs.

The iscript select cdna synthesis kit is a sensitive, flexible, and easytouse kit for the generation of firststrand cdna. I will be carrying out a twostep qrtpcr, which involve the first step of first strand cdna synthesis. The monsterscript 1ststrand cdna synthesis kit has been discontinued. Cholesterol and 27hydroxycholesterol promote thyroid. It can finalize first strand cdna synthesis in just 30 minutes, which reduces the total time required for rtpcr. The transcriptor universal cdna master is a convenient fast solution for cdna synthesis and realtime pcr analysis. Quantification of the g6pdh gene with the transcriptor first strand cdna synthesis kit and the lightcycler 2. Since there is no need to perform a separate dnase digestion and. Betaine reduces pausing and stops by the reverse transcriptase, enabling improved reverse transcription through difficult sequences, such as regions of high gc content. Thermo scientific revertaid first strand cdna synthesis kit is a complete system for efficient synthesis of first strand cdna from rna templates. Firststrand cdna synthesis for quantitative rtpcr origene. Easy and reliable qrtpcr 1 introduction analysis of total. 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.

The transcriptor first strand cdna synthesis kit is designed to reverse transcribe rna mrna, total rna, viral rna, and in vitro transcribed rna from a variety of sources for the following applications. Accuscript high fidelity 1st strand cdna synthesis kit. Study gene expression levels, via twostep rtpcr, using qualitative rtpcr on. However, amplification of some pcr targets 1 kb may require the removal of rna complementary to the cdna. Prepare a cdna synthesis master mix according to one of the tables below, depending on the cell lysate volume. 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. The transcriptor high fidelity cdna synthesis kit provides all reagents required for first strand cdna synthesis reactions. Using the transcriptor first strand cdna synthesis kit, total rna was reverse transcribed by random hexamer priming. Transcriptor reverse transcriptase the core component of the kit is a recombinant reverse transcriptase expressed in e. Transcriptor first strand cdna synthesis kithigh pure kit.

First strand cdna synthesis after thawing, mix and briefly centrifuge the components of the kit. Reverse transcription pcr rtpcr was performed using a transcriptor first strand cdna synthesis kit roche, basel, switzerland and tagged primers see table 1 for sequences to add a specific. First strand cdna from roche, goscript, high capacity cdna. First strand cdna synthesis kit direct pcr reagents and. Two cdna synthesis primers are provided with the kit. Transcriptor firststrand cdna synthesis kit roche diagnostics mannheim.