DescriptionTo complement the SensiFAST™ Probe and SYBR® kits, Meridian has developed the SensiFAST cDNA Synthesis Kit.Product HighlightsEfficient – high-target affinity, coupled with a novel TransAmp buffer system for improved yield of full-length cDNAUnbiased – optimized mix of random hexamers and anchored oligo dT primers for complete 5’ to 3’ RNA sequence representationSensitive –lower Ct values from a broad range of input cDNA concentrations, enabling accurate detection of very low-copy targetsRobust – reliable reverse transcription under challenging conditions, including complex templates and in the presence of inhibitorsFast – high-yield reverse transcription from a very broad of targets in as little as 5 minutesProduct DescriptionThe SensiFAST™ cDNA Synthesis Kit provides a rapid and sensitive method for first-strand cDNA synthesis, which displays excellent linearity across a wide range of starting material. This gives the same relative representation in cDNA templates, regardless of gene abundance, making it excellent for use in qPCR studies.SensiFAST cDNA Synthesis Kit contains a highly-pure reverse transcriptase and optimized TransAmp™ buffer system, which includes a unique blend of random hexamers and anchored oligo (dT) primers to deliver the highest quality qPCR ready cDNA. This makes the SensiFAST cDNA Synthesis Kit ideal for working with limited sample volumes, such as laser-micro dissected samples and tissue biopsies (down to 1 pg of input RNA), to reverse transcribe precious RNA into stable cDNA ready for accurate real-time quantification. The unique blend of random hexamer primers and anchored oligo dT in the TransAmp Buffer also ensure unbiased 3’ and 5’ coverage and reverse transcription of all regions.SensiFAST cDNA Synthesis Kit can be used with SensiFAST Probe and SensiFAST SYBR® Kits for fast real-time RT-qPCR without compromising on quality, giving real-time results in less than an hour.Request InfoApplicationsGene expression analysisTissue biopsy analysismiRNA profiling / quantificationRNA target detectionPathogen detection Fig. 1 Speed and sensitivitySensiFAST cDNA Synthesis Kit and a kit from supplier B were used for reverse transcription of a specific target from total RNA under fast conditions. The results illustrate that the SensiFAST cDNA Synthesis Kit (red) is more sensitive than the kit from supplier B (blue), as judged by the lower Ct values. Fig. 2 Reproducibility of SensiFAST cDNA Synthesis KitSensiFAST cDNA Synthesis Kit was employed in 48 independent first-strand reactions, containing 100 ng total RNA and another 48 reactions containing 100 pg. The resultant first-strand product from each RT reaction was used in triplicate qPCR assays. The results demonstrate the excellent reproducibility of SensiFAST cDNA Synthesis Kit at high and low input RNA, as judged by the same Ct values. Fig. 3 Unbiased representation across target genesSensiFAST cDNA Synthesis Kit and a kit from supplier B were for reverse transcription of a specific target from total RNA. Primer pairs were then designed against the transcript at 1 kb intervals and used in qPCR reaction with SensiFAST SYBR® No-ROX. The consistent Ct values show that, unlike the results from supplier B (blue), the SensiFAST cDNA Synthesis Kit (red) did not show any bias across the target. Fig. 4 Speed of SensiFAST cDNA KitSensiFAST cDNA Synthesis Kit was used for reverse transcription of a specific target from a 10-fold serial dilution of total RNA, for either 5 min (blue) or 60 min (red) at 42 °C. The results illustrate the high efficiency of reverse transcription over a broad linear dynamic range under both standard and fast conditions. SensiFAST cDNA Synthesis Kit Customer Review SensiFAST cDNA Synthesis Kit Customer ReviewReal-Time PCR Selection ChartOne-step Vs. Two-step real-time RT PCR A discussion of the pros and cons of each detection strategy.