Plant-cDNA-Genemed Synthesis Inc.

Abstract The quantity and quality of first-strand cDNA directly influence the accuracy of transcriptional analysis and quantification. Using a plant-derived α-tubulin as a model system, the effect of oligo sequence and DTT on the quality and quantity of first-strand cDNA synthesis was assessed via a combination of semi-quantitative PCR and real-time PCR. The results indicated that anchored oligo dT significantly improved the quantity and quality of α-tubulin cDNA compared to the conventional oligo dT. Similarly, omitting DTT from the first-strand cDNA synthesis also enhanced the levels of transcript. This is the first time that a comparative analysis has been undertaken for a plant system and it shows conclusively that small changes to current protocols can have very significant impact on transcript analysis. Key words cDNA; real-time PCR; RT-PCR; transcript analysis In the past 10 years, there was a very rapid develop-ment in fluorescence-based techniques for the detection and monitoring of transcript abundance. Two methods that have gained rapid popularity are microarray and real-time PCR. The microarray allows parallel analysis of thousands of genes from two differentially labelled RNA populations

In vitro destabilization of plant viruses and cDNA synthesis

Plant Direct; Blood Direct; RT-PCR – cDNA synthesis

What is the best method for cDNA synthesis? - …

Substantial progress has been made in recent years in the cloning of cDNAs encoding the proline biosynthetic enzymes from higher plants by complementation of proline-requiring mutants of (Verma et al, 1992).

Plant Tissue cDNA : Zyagen, Life Science Products

Annatto is one of the important natural food colourant widely used in dairy industry. Quality RNA in large quantity is often required in the analysis of gene expression. RNA extraction from samples collected from woody plants is generally complex, and becomes the main limitation to study gene expression, particularly in crops like Bixa orellana. Standard RNA extraction protocols are time consuming, laborious and cannot be adapted for high throughput functional analysis. Therefore a simple and effective protocol for extraction of high quality total RNA from bark tissue of woody stem was achieved using the RNeasy plant mini kit (Qiagen, USA). The extracted RNA was successfully converted into double-stranded cDNA using the SMATer cDNA synthesis kit (Clontech, USA) which is based on the Switching Mechanism At 5’ End of RNA Transcript (SMART) technology. The integrity of the total RNA used for synthesizing double stranded cDNA was assessed agarose gel electrophoresis and by PCR. As expected, the PCR product contained the full coding sequence plus 69 and 196 bp of 5’ and 3’ UTRs respectively. The double-stranded cDNA was used successfully for creating a SSH cDNA library. The cDNA could also be useful for a number of other applications like cDNA library construction, EST analysis, RACE and Next Generation Sequencing (NGS).

14/03/2012 · A Cost-effective Double-Stranded cDNA Synthesis for ..
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cDNA Synthesis & Library Construction - Clontech

Total RNA used for cDNA synthesis is isolated by modified guanidine thiocyanate techniques

Pesquise com mais rapidez e eficiência aqui! Cdna Synthesis

RACE (Rapid Amplification of cDNA Ends) 5 Prime RACE and 3 Prime Overview

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