Applied Biosystems™ TaqMan™ Advanced miRNA cDNA Synthesis Kit

Applied Biosystems™ TaqMan™ Advanced miRNA cDNA Synthesis Kit

The Applied Biosystems™ TaqMan™ Advanced miRNA cDNA Synthesis Kit uses a universal reverse transcription (RT) chemistry to prepare the cDNA template for use with TaqMan™ Advanced miRNA Assays for detection and quantification of mature miRNAs in biological samples.

The TaqMan Advanced miRNA cDNA Synthesis Kit uses 3' poly-A tailing and 5' ligation of an adaptor sequence to extend the mature miRNAs present in the sample on each end prior to reverse transcription. Universal RT primers recognize the universal sequences present on both the 5' and 3' extended ends of the mature miRNAs. All mature miRNAs in the sample are reverse transcribed to cDNA.

Features

• Sensitivity: high sensitivity allows for quantification of low-expressing miRNA targets, especially in biological fluids such as plasma, serum, and tissue
• Flexibility: the universal RT chemistry generates universal cDNA templates for all miRNAs present in the sample, allowing for flexibility in study design
• Sample savings: requires only 2 μL of plasma/serum sample input and generates enough cDNA for 600 qPCR reactions (20 μL) or 1200 qPCR reactions (10 μL)
• Specificity: 5’ ligation step allows for single base pair discrimination along entire miRNA sequence, including difficult-to-detect mismatches on 5’ end
• Gold-standard TaqMan chemistry: This kit is designed for use with TaqMan® Advanced miRNA Assays for qPCR, which draw on Thermo Fisher Scientific's industry-leading bioinformatics assay design pipeline and validation system to help ensure high specificity and minimal cross-reactivity, even for closely-related miRNA families.

Contents

• 10X PolyA Buffer
• ATP, 10mM
• Poly A Enzyme, 5 U/μL
• 5X Ligase Buffer
• RNA Ligase, 10 U/μL
• 50% PEG 8000
• 25X Ligation Adaptor
• 10X RT Enzyme Mix
• 20X Universal RT Primer
• dNTP Mix, 100mM
• 20X miR-Amp Primer Mix
• 5X RT Buffer
• 2X miR-Amp Master Mix