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. It can be used with either sequencespecific primers, polydt15 primers, or. Rneasy protect mini kit for immediate stabilization of rna in harvested animal tissues and subsequent total rna purification. Omniscript reverse transcriptase for firststrand cdna synthesis twotube rtpcr pdf 381kb english format file size language download get adobe reader contact qiagen. 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. The accupower rt premix is a master mix for cdna synthesis that consists of an easy to resuspend, lyophilized mix of mmlv reverse transcriptase, rnase inhibitor, dntps, reaction buffer, tracking dye, and patented stabilizer. Rnaquant cdna reaction setup for 1 rna sample to be assayed on qpcr 96well plates it is important to start with total rna that includes the lncrna fraction. Firststrand cdna synthesis kit ge healthcare, 27926101. 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. Choose the best cdna synthesis kit for your application, with formulations optimized for ease, yield, flexibility, fidelity, or gdna clearance. For cdna synthesis with integrated removal of genomic. These are available online in convenient and compact pdf format at. For priming, three different primer systems can be used.
Highcapacity cdna reverse transcription kits product. Mix thoroughly by pipetting up and down several times. The latter is designed to bind at the beginning of the polya tail to generate fulllength. The enzymes lack of rnase h activity contributes to its ability to make longer cdnas and more complete fulllength libraries of firststrand cdna. However, this step can be omitted in some cases unpublished results. In rtpcr, after an initial reverse transcription step to produce the cdna template, pcr is performed to amplify the target sequence. Once the sequence of the forward primer has been determined, the primer should be ordered through a custom oligo supplier. Mmlv cdna synthesis kit provides a sensitive and easytouse solution for twostep rtpcr. The qscript cdna synthesis kit is a sensitive and easytouse solution for rna. 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.
I want to to do pcr for a specific gene of interest. Tetro cdna synthesis kit contains our highly sensitive mmlv reverse transcriptase, oligo dt 18 and random hexamer primers and all the necessary components to generate high quality cdna from rna templates fig. After completion of cdna synthesis, use 12 l of the first strand reaction out of the 20 l for pcr amplification. The user can use their own gene specific primers or either the randomers or oligodt included separately in the kit. Omniscript reverse transcription handbook en qiagen. The master mix kit is used for first strand cdna synthesis from rna. 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. The kit uses mmlv reverse transcriptase with lower rnase h activity, which avoids degradation of rna when rnadna hybrid forms in. Since there is no need to perform a separate dnase digestion and. Random random primers ensure that the first strand synthesis occurs efficiently with all species of rna molecules present, including mrna and rrna. I have purified rna and synthesized cdna using quantitech reverse transcriptase kit from qiazen. The firststrand cdna generated is ideal for pcr fig. Includes goscript reverse transcriptase, mmlv and amv reverse transcriptases. The preblended 5x iscript reverse transcription supermix contains all the necessary components, except rna template, in one tube for firststrand cdna synthesis.
Make a cdna synthesis reaction mix by adding the iscript reverse transcription supermix to the dnasetreated rna template according to the guidelines in table 4. Firststrand cdna synthesis kit is designed to generate fulllength firststrand cdna from mrna templates. Teloprime fulllength cdna amplification kit v2 lexogen. Quanta qscript cdna synthesis kit quanta qscript cdna supermix qiagen quantitect reverse transcription kit stratagene affinityscript qpcr cdna synthesis kit. Allow the primer to anneal to the template for 10 minutes at room temperature. The mirna 1ststrand cdna synthesis kit can accommodate a range of total rna input amounts from 30 ng to 1. Ma255e messagebooster cdna synthesis kit for qpcr 72017. Genetics and molecular biology reverse transcriptase and cdna synthesis kunitada shimotohno encyclopedia of life support systems eolss 1. The rnaquant cdna synthesis kit prepares any rna for qpcr the polya tailing and tagging method in the rnaquant system generates cdna that is highly unique and, when used with the 3 universal reverse primer, does not produce any signal in the absence of reverse transcriptase or lead to the detection of any residual genomic dna. For cdna synthesis and genomic dna elimination in rna samples for use with rt 2. The 10x superscript enzyme mix includes superscript iii rt. Transcriptase mmlv rt is an rnadependent dna polymerase that can be used in cdna synthesis.
The optimized 5x transamp buffer includes all necessary components for cdna synthesis, including a unique blend of oligo dt and random hexamer primers and the ribosafe rnase inhibitor. The revertaid first strand cdna synthesis kit is supplied with both oligodt 18 and random hexamer primers. This kit includes five tubes comprehensive of the reagents required for successful rtpcr. Efficiency kits varied in their ability to efficiently generate cdna from different rna inputs linearity of cdna synthesis kits with bactin 15 20 25 30 35 401. Messagebooster cdna synthesis kit for qpcr lucigen. The oligodt 18 primer anneals selectively to the polya tail of mrna. Invitrogen superscript vilo cdna synthesis kit life. The iscript gdna clear cdna synthesis kit is a sensitive, fast, and convenient reagent for gene expression analysis and realtime quantitative pcr qpcr. The adapter primer has a unique sequence at its 5 end.
For reliable first strand cdna synthesis from all rna sources. Rtpcr kits and standalone reverse transcriptase enzymes for reverse transcription of fulllength cdna from your experimental sample. Doublestranded dna can be used to make nondirectional cdna libraries, to enrich differentiallyexpressed. Thermo scientific verso cdna synthesis kit provides robust transcription of rna to create a complete cdna pool. Rneasy mini kit for purification of total rna from animal cells, animal tissues, bacteria, and yeast, and for rna cleanup. The user can use their own gene specific primers or either the randomers or oligodt included separately in. Universal rt primers recognize the universal sequences present on both the 5 and 3 extended ends of the mature mirnas.
The resulted cdna is singlestranded and this process is called reverse transcription rt or first strand cdna synthesis. Following firststrand cdna synthesis, the sample can be used directly for in vitro amplification using pcr or for secondstrand synthesis using the gublerhoffman method. Revertaid first strand cdna synthesis kit thermo fisher scientific. The sensifast cdna synthesis kit displays excellent linearity across a wide range of starting material, revealing the same relative representation in cdna templates, regardless of gene abundance, making it excellent for use in realtime pcr studies. Random hexamer primers do not require the presence of the polya tail, therefore, they can be used for transcription of the 5end regions of mrna or cdna. To apply pcr to the study of rna, the rna sample must first be reverse transcribed to cdna to provide the necessary dna template for subsequent pcr. Productfinder helps you to find the best qiagen product for your application. With the qscript microrna cdna synthesis kit mirnas are polyadenylated in a polya polymerase reaction. Downstream applications include cdna amplification with reverse transcription pcr. 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. The monsterscript 1ststrand cdna synthesis kit has been discontinued. The teloprime fulllength cdna amplification kit v2 is a protocol for generating fulllength cdna from total rna. In contrast to kits from other suppliers, the quantitect reverse transcription kit provides high yields of cdna template for realtime pcr.
It can be used with either sequencespecific primers, polydt15 primers, or random primers, pdn6. This step is very important in order to perform pcr since dna polymerase can act only on dna templates. The kit provides all reagents required for firststrand cdna synthesis reactions. The kit can provide high efficient polya modification and reverse transcription and efficiently synthesize the firststrand cdna from 20 pg2. For purification of up to 10 mg endotoxinfree advanced transfectiongrade plasmid or cosmid dna. Introduction two groups working independently in the united states in 1970 discovered reverse transcriptase activity in retroviruses. Hi, in our lab we study gene expression in plants and good rna and cdna synthesis is very important.
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. Omniscript reverse transcriptase for firststrand cdna synthesis twotube rtpcr pdf 381kb english format. The reversetranscription master mix contains all components required for first strand cdna synthesis except template rna. Two cdna synthesis primers are provided with the kit. This gives the same relative representation in cdna templates, regardless of gene abundance, making it excellent for use in qpcr studies. The sensifast cdna synthesis kit provides a rapid and sensitive method for firststrand cdna synthesis, which displays excellent linearity across a wide range of starting material. Kit components include premixed oligonucleotides and reverse transcriptase, which facilitate applications like gene expression, rna quantification, cloning, and sequencing. This is recommended when the cdna synthesis reaction uses less than 10 ng of rna input. The tetro cdna synthesis kit is the more flexible solution. Some couple cdna synthesis and quantitative realtime pcr in a single reaction. Make your selections and you will find the most suitable products. Monsterscript reverse transcriptase is a thermostable reverse transcriptase that completely lacks rnase h activity. Mint2 cdna synthesis kit contains different plugoligo and cds adapters allowing synthesis of cdna with different flanking sequences. Mmlv reverse transcriptase 1ststrand cdna synthesis kit lucigen.
The rt primer mix contains a specially optimized mix of oligodt and random primers that enable cdna synthesis from all regions of rna transcripts, even from 5 regions see figure sensitive detection of a target at the 5 region of a 12. The applied biosystems highcapacity cdna reverse transcription kit uses the random primer scheme for initiating cdna synthesis. The tonkbio first chain cdna synthesis kit for rtpcr is optimized to efficiently synthesize first chain cdna from mrna or total rna templates. The mmlv hreverse transcriptase is optimized for reliable cdna synthesis over a wide dynamic range of input rna. Unesco eolss sample chapters genetics and molecular biology reverse transcriptase and cdna synthesis kunitada shimotohno encyclopedia of life support systems eolss 3. Protocol for cdna synthesis and qrtpcr cdna synthesis. Store the remaining first strand cdna sample at 20 c until use. Please refer to the section below to choose the pair of adapters suitable for your needs. You selects the desired setup, especially the desired rt primer. 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 enzyme is exceptionally well with a wide variety of targets. Rneasy plant mini kit for purification of total rna from plants and filamentous fungi.
This step uses the qiagen rneasy minelute cleanup kit qiagen cat. The first strand cdna synthesis kit is used for the synthesis of the first strand cdna as the starting reaction for rtpcr. Readytouse master mix for cdna synthesis from total rna. January 2017 for the synthesis of doublestranded cdna from total rna or mrna cat. Using the superscript vilo cdna synthesis kit this formulation provides enhanced cdna synthesis efficiency and can be used with very low and very high amounts of input rna up to 2. 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.1023 792 399 1584 48 659 167 625 1016 908 689 647 1324 1450 154 313 547 1432 1261 71 694 685 1130 1575 67 923 1624 953 1497 422 996 168 878 1114 1046 1215 756 685 1231 1284 1112 601