The first strand cdna synthesis kit allows the user to choose the desired primer fur cdna synthesis. Reactions can be scaled up to 100l to generate 10g of cdna from single reaction. Rtpcr cdna synthesis kits thermo fisher scientific. A twostep rt pcr format is useful for amplifying multiple targets from a single. Smart technology is especially useful for researchers who have limited starting material, such as total rna from a small sample.
Smarter pico pcr cdna synthesis kit user manual takara bio. Bd advantage 2 pcr enzyme system the wolfson centre for. Guide to using the smarter pico cdna synthesis protocol for pcrselect cdna. 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. Pr782357 17 smart race cdna amplification kit user manual. Formaldehyde agarose gel visualization with ethidium bromide etbr. A twostep rtpcr format is useful for amplifying multiple targets from a single.
Energicscript first strand cdna synthesis kits for qpcr user. Refer to nanodrop protocol for instructions on use. 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. For greater application flexibility, hexamer primers, allowing all rnas in the reaction to be used as templates, and an oligo dt primer, for the synthesis of cdna from only polya tailed mrna, are. Affinityscript multiple temperature cdna synthesis kit. The superscript iii cellsdirect cdna synthesis kit is optimized for synthesizing firststrand cdna directly from a mammalian cell lysate without first isolating the rna. Pcr cdna synthesis kit user manual united statescanada 800. Superscript iv rt is very robust and efficient and can deliver up to 100x higher cdna yields with degraded rna than other commercially available rts figure 1. The amplified cdna populations under comparison are then subjected to suppression subtractive hybridization sshpcr, a technique that couples subtractive hybridization with suppression pcr to selectively amplify fragments of differentially expressed genes. Higher yields and precision than other cdna synthesis kits at a fraction of the cost.
The 10x superscript enzyme mix includes superscript iii rt. The smarter cdna synthesis method is also optimized for rapid amplification of cdna ends race. Clontechs smart cdna synthesis kit biocompare product. Smarter pico pcr cdna synthesis kit user manual 020916. The product of the first strand cdna synthesis can be used directly in pcr or qpcr. The qpcrbio cdna synthesis kit uses the latest developments in reverse transcriptase technology and buffer chemistry to enhance cdna synthesis speed and yield with accurate transcript representation. Some couple cdna synthesis and quantitative realtime pcr in a single reaction. High quality rna preparation is critical for the synthesis of cdna for pcr. Kit components and storage conditions upon receiving the teloprime kit, remove the smaller inner box and store it in a 20 c freezer. Neb offers several reagents for cdna synthesis for use in applications including qpcr and qrtpcr. The kit amplifies the mrna contained in a small total rna sample and then converts the amplified rna to cdna that is ready for pcr or qpcr fig. Two cdna synthesis primers are provided with the kit. Smart technology is used in a ligationfree workflow to generate sequencing libraries for illumina platforms.
Synthesis kit and manual agarosegel size selection. Apr 06, 2000 thats not to say that the smart cdna synthesis kit is perfect. Invitrogen superscript iv rt, the most recently introduced enzyme, was engineered to offer superior cdna synthesis performance with even the most challenging rna samples. Purchase of the bd smart race cdna amplification kit includes a free sample of bd powerscript reverse transcriptase and a free trialsize bd advantage 2 pcr kit cat. Monsterscript reverse transcriptase is a thermostable reverse transcriptase that completely lacks rnase h activity. Not recommended or intended for diagnosis of disease in humans or animals. Isoseq using the clontech smarter pcr cdna synthesis kit. The combination of highly efficient cdna synthesis, effective rnase inhibition and pure dntps allows high yields of cdnas of more than 19 kb. Please note that there are reagents in this kit that might not be needed when used in conjunction with the generacer kit.
Summary the smarter pico pcr cdna synthesis kit provides a pcrbased method for producing highquality cdna from picogram quantities of total rna. Highcapacity rnatocdna kit thermo fisher scientific. I am planning to do rt pcr on rna samples 5ug which are quite precious because they are not mine, they were a gift. C, ssh see clontech smart pcr cdna synthesis kit user.
The incorporation of bd smart technology also permits. General considerations for smart race amplification 11 v. Bd smart race cdna amplification kit user manual i. For cdna synthesis and genomic dna elimination in rna samples for use with rt 2. The volume of first strand cdna synthesis reaction mixture should not comprise more than 110 of the total pcr reaction volume. To use the zero blunt topo pcr cloning kit for sequencing, please refer to the manual supplied with this kit. When using this kit, beware of the included chromaspin columns for purification of your cdna. Therefore, this kit is best used with intact total rna preparations. Verso 1step rtpcr kit, with thermostart taq hot start ab.
The integrity of total rna can be visually assessed by the ratio of 28s. The cornerstone of the smarter race cdna amplification kit is smart technology, which eliminates the need for problematic adaptor ligation and lets you use firststrand cdna directly in race pcr, a benefit that makes race far less complex and much faster chenchik et al. Lysis and reverse transcription are performed in the same tube, and the resulting firststrand cdna is ready to use in cloning and pcr. Energicscript first strand cdna synthesis kits user manual description the energicscript cdna synthesis kit includes all the necessary reagents for cdna synthesis to be used in qpcr.
Infusion smarter directional cdna library construction. Applications such as northern blot analysis, cdna library construction and cdna subtraction are demanding when it comes to starting material. If it is just for realtime pcr, random primers will be fine and this will not be. Either total rna, mrna, viral rna or in vitro transcribed rna can be used as a template for reverse transcription. Invitrogen superscript vilo cdna synthesis kit life. Applied biosystems highcapacity cdna reverse transcription kit. Using one of the required thermal cyclers listed in the highcapacity rnato cdna kit user guide pub. Do not use internally or externally in humans or animals.
The kit is an optimal choice for generating high quality cdna from viral rna, mirna or other targets for qpcr or for pcr. Materials and kits needed preparing cdna from rna samples firststrand synthesis first strand cdna synthesis employs the clontech smarter pcr cdna synthesis kit. Schematic of technology used by the smarter smrnaseq kit for illumina. The rest of the kit components should be stored at room temperature and protected from light. Upscaling cdna synthesis using superscript iii reverse. Together, these products provide sufficient reagents for 7 cdna synthesis reactions and 30 pcr reactions. I am planning to do rtpcr on rna samples 5ug which are quite precious because they are not mine, they were a gift. For your convenience, reagents are available as kits or standalone products, depending on your needs. Before use, check the contents of cb1, cb2, and cw. For the maximum convenience the kit includes even the pcr water to set up the reaction, so the only thing you need to take care of is the high quality rna template. For the cdna synthesis, i am going to use superscript iii rt with oligo dts. Bd smart cdna library construction, bd smart pcr cdna synthesis. This effect is even observed for long mrna templates and low concentrations of total rna 25 ng.
The kit uses a linear rna amplification process to amplify the rna from a small population of cells, then converts the amplified rna to cdna that is ready for pcr. Using a water bath or thermal cycler for this incubation may reduce the volume of the reaction mixture due to evaporation, and therefore reduce the ef ficiency of firststrand synthesis. Smarter race 53 kit user manual 052617 takara bio usa, inc. 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. The use of this cdna synthesis procedure, in combination with t4 gene 32 protein, increases the yield of rtpcr products from approximately 90% to 150%. This kit saves time and reduces contamination due to a. The 50reaction pack size also includes 10 control reactions control rna and control primer mix.
Introduction continued the smart ii a oligonucleotide and 3 smart cds primer ii a provided in the smart pcr cdna synthesis kit each have an rsa i site to facilitate removal of these identical sequences from the pcramplified cdna molecules. Transcriptor high fidelity cdna synthesis kit accuracy. Neb offers several reagents for cdna synthesis for use in applications including qpcr and qrt pcr. Hi, in our lab we study gene expression in plants and good rna and cdna synthesis is very important. Superscript iv reverse transcriptase thermo fisher.
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. This user manual includes two protocols for cdna synthesis. 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. The one step rt pcr kit provides an efficient, accurate, and convenient way to synthesize cdna and amplify via pcr, from any template, compatible with both polya mrna and total rna. 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. Unless size exclusion is an absolute necessity, much better yields can be achieved with a standard pcr purification method such as clontechs nucleospin kit or qiagens. Usb firststrand cdna synthesis kit for realtime pcr product number 75780 50 reaction size 20.
Pcr primer sequences the table below lists the sequence of the pcr primers included in the generacer kit. Notice to purchaser advantage 2 products are covered by u. Quanta qscript cdna synthesis kit quanta qscript cdna supermix qiagen quantitect reverse transcription kit stratagene affinityscript qpcr cdna synthesis kit. Smarter pcr cdna synthesis kit user manual takarabio. This kit saves time and reduces contamination due to a reduced number of pipetting steps required for rt pcr set up. The cornerstone of smarter cdna synthesis is still our unique smart switching mechanism at the 5 end of rna transcript technology. Mint cdna synthesis kit is designed to synthesize full length enriched. Affinityscript multiple temperature cdna synthesis kit 3 firststrand cdna synthesis protocol notes wear gloves at all times during the firststrand cdna synthesis and pcr amplification procedures and while handling materials and equipment to prevent contamination by ribonucleases rnases. Invitrogen superscript iii cellsdirect cdna synthesis kit. I recommend you to use super script kit for cdna synthesis. Smarter smrnaseq kit for illumina user manual 121219 takara bio usa, inc.
Messagebooster cdna synthesis kit for qpcr gene target. The monsterscript 1ststrand cdna synthesis kit has been discontinued. Protocol for cdna synthesis and qrtpcr cdna synthesis. Home shop all products kits and assays realtime pcr and endpoint rtpcr kits. Efficiencies of cdna synthesis measured in twostep qrtpcr choosing the best qrtpcr product for your application o ne step vs two step. The smarter pico pcr cdna synthesis kit provides a pcrbased method for producing highquality cdna from picogram quantities of total rna. The invitrogen superscript line of reverse transcriptases rts represents the most widely used and highly referenced rts for cdna synthesis in many applications. These conditions are optimized for use with the highcapacity rnato cdna kit. Overview of the messagebooster cdna synthesis kit procedure.
The reverse transcriptase, buffer system and combination of random hexamers with anchored oligodt allow for unbiased, efficient, sensitive cdna. Smart race cdna amplification kit user manual table of contents i. Superscript iv rt is a superior choice for cdna synthesis with any type of rna and. Reverse transcriptase, which has lower rnase h acti first. An efficient rt will reversetranscribe even the most difficult types of rna, including those that are of low abundance or degraded. Downstream applications include realtime pcr, standard pcr, and microarrays. Get better results with smarter pcr cdna synthesis kits. For priming, three different primer systems can be used.