Superscript iv first strand synthesis kit invitrogenis. Add reaction mix to annealed rna rt reaction setup use the measurements below to prepare your rt reaction, or enter your own parameters in the column provided. Dnafree total rna from jurkat t cellleukemia line and mrc5. Reverse transcription of rna into cdna protocol osski. Superscript iii firststrand synthesis system for rtpcr author. The protocol described in this article provides a classic and simple means to generate 3. Increase the temperature of firststrand reaction up to 55c. System for cdna synthesis, invitrogen life technologies, pn 18090019. Extralong firststrand cdna synthesis future science. Prior to cdna synthesis, please make sure that your rna is intact and free of. Superscript ii reverse transcriptase thermo fisher scientific. Superscript vilo cdna synthesis kit product information. The 260280 and 260230 values of my rna samples are above 2. Purified rna must be free of salts, metal ions, ethanol and phenol to avoid inhibiting the cdna synthesis reaction.
Protocols cdna synthesis from total rna or mrna since the conditions described for the first strand reaction are optimized, do not change volumes and concentrations. This formulation provides enhanced cdna synthesis efficiency and can be. Alternative protocol for onecycle cdna synthesis from total rna. Mix rna sample and primer dt 23 vn in a sterile rnase free. Maintain an elevated temperature after the annealing step, as described in the protocol for cdna synthesis from highgc content transcripts, page 3. Reaction protocol incubate the complete reaction mix in a thermal cycler using. Components of thermo scientific revertaid first strand cdna synthesis kit.
C recommendation for optimal results using storage and stability the iscript cdna synthesis kit the maximum amount of the cdna reaction that is. Firststrand cdna synthesis using superscript ii rt. Race system for rapid amplification of cdna ends cdna synthesis directly from cells using superscript iii cellsdirect cdna synthesis system cdna synthesis from transcripts with high secondary structure using thermox reverse transcriptase superscript iii firststrand synthesis supermix. Protocol for cdna synthesis and qrtpcr cdna synthesis. The cdna synthesis system can be used to produce doublestranded cdna by two procedures.
The smarter pcr cdna synthesis kit is an improved version of. The invitrogen superscript vilo cdna synthesis kit provides the high temperature capability of superscript iii reverse transcriptase in an optimized format for generating firststrand cdna for use in realtime quantitative rtpcr qrtpcr. Thaw all necessary components and place them on ice. Guide to using the smarter cdna synthesis protocol for pcrselect cdna.
345 1110 719 47 1159 1327 454 1488 575 1184 501 235 1127 1289 1403 1383 362 646 1030 455 695 1173 609 1272 237 946 1282 1254 537 192 1159 1302 1399 84 637 958