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iScript cDNA Synthesis Kit | Life Science Research | Bio-Rad

cDNA Synthesis | Bio-Rad

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Bio-Rad's iScript cDNA Synthesis Kit | Biocompare …

This protocol is adapted from the following sources:cDNA/AFLP ProtocolPreparation of Para-magnetic beads from Promega cat#Z5482:a) suspend magnetic particles in bottle - transfer 200ul (200 ug) of beads per sampleof RNA to be purified to a microfuge tube or tubes.b) place microfuge tube in magnetic stand for 30 sec or longerto collect the beads.c) remove supernatent and wash 3X with 200 ul of 0.5XSSCd) finally resuspend the SA-PMPs in 100ul of 0.5X SSCfor each 200 ul of beads originally removed - each 100 ulaliquot should be in its own labelled microfuge tube, 1 for eachRNA sample.note: this should not be done more than 30 mins before thebeads are to be used. Hybridization of biotinylated polyT primer:a) add H2O to 20 ug - 200ug (more is better) ofRNA to make a final volume of 500 ulb) heat to 65C for 10 mins.c) add 1.5 ul of 100 uM poly d(T)25V and 13ul of 20X SSCd) let cool to room temperature (~10min)e) add RNA/poly T mix to the previously prepared 100 ul ofSA-PMPs; mix gently and let sit at room temperature for 10minf) capture PMPs with magnetic stand - carefully removesupernatent and wash 3X with 300 ul of 0.1X SSC.g) wash w/ 50 ul of 1X reverse transcriptase buffer. 1rst Strand cDNA synthesis:a) aliquot samples of Reverse Transcriptase Mix:b) heat mix to 37C for 2 mins - then add 1 ul (50U)of MMLV reverse transcriptase (Stratagene).c) incubate at 37C for 15 mins, then 42C for 45mins, mixingevery 15mins2nd Strand synthesis:a) add 140 ul of 2nd strand cDNA rxn mix to 1rststrand synthesis rxn:

03/01/2006 · A review of the Bio-Rad's iScript cDNA Synthesis Kit

ProtoScript II – NEB ProtoScript II First Strand cDNA Synthesis Kit and random primer mix; SuperScript II – Invitrogen SuperScript First Strand Synthesis System for RT-PCR and random hexamers; SuperScript® III – Invitrogen SuperScript III First Strand Synthesis SuperMix and random hexamers. The following reagents are supplied with this product:

iScript™ cDNA Synthesis Kit from Bio-Rad | SelectScience

Read independent reviews on iScript™ cDNA Synthesis Kit from Bio-Rad on SelectScience

With an optimized section of primers, this kit isappropriatefor RT-PCR, real-time RT-PCR, and the synthesis of longer cDNAfragmentsfor applications such as cloning.

The RNA was reverse transcribed using first strand cDNA synthesis kits, as indicated, and 1/10th of the cDNA product was used for qPCR using luciferase-specific primers and iTaqTM Universal SYBR Green Supermix (BioRad).

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The PrimeScript First Strand cDNA Synthesis Kit contains all of the reagents necessary for synthesis of first-strand cDNA from total or poly(A)+ RNA using PrimeScript Reverse Transcriptase. Choose this cDNA synthesis kit when you want to synthesize full-length cDNA with provided primers (select from oligo dT or random 6-mer primers, both included in the kit.) This cDNA synthesis kit is powered by PrimeScript Reverse Transcriptase, which has exceptionally strong strand displacement activity and efficiently synthesizes cDNA. PrimeScript RTase is robust, versatile and well-suited for applications requiring full-length cDNA such as cDNA library construction. This PrimeScript-based cDNA synthesis kit also works well with other cDNA-dependent techniques including RT-PCR, cDNA probe preparation and real-time quantitative RT-PCR (qRT-PCR or RT-qPCR). The PrimeScript First Strand cDNA Synthesis Kit is ideal for reverse transcription of many different RNA templates, including GC-rich templates and RNAs with high levels of secondary structure.

cDNA synthesis and PCR amplification steps are performed ina single reaction using gene-specific primers, resulting in a streamlined RT-PCRprotocol.

The kit combines three optimized mixes: OneTaq One-Step Enzyme Mix,OneTaq One-Step Reaction Mix and OneTaq One-Step Quick-Load ReactionMix.

PrimeScript 1st strand cDNA Synthesis Kit - Clontech
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    iScript Select cDNA Synthesis Kit | Bio-Rad Laboratories, Inc.

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iScript Select cDNA Synthesis Kit ..

First introduced in Japan, PrimeScript is now available to scientists worldwide. Over 3,700 peer-reviewed articles have been published in which PrimeScript Reverse Transcriptase was referenced in a variety of scientific applications such as gene expression, gene discovery, transcriptome analysis, miRNA expression and regulation, molecular evolution, virology and microbiology. PrimeScript RTase is a modified, recombinant MMLV (Moloney Murine Leukemia Virus) reverse transcriptase and is verified to have low levels of RNase H. Because of the excellent extension capability of PrimeScript RTase, cDNA synthesis reactions can be performed at a lower temperature (42°C), decreasing the risk of RNA degradation that can occur during conventional reactions performed at higher temperatures.

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