Custom RT2 lncRNA PreAMP Primer Mix
For cDNA synthesis prior to real-time PCR
Analysis of small amounts of RNA is critical for biopsy, FFPE, and other types of samples. RT2 lncRNA PreAMP Primer Mixes enable lncRNA expression analysis from as little as 1 ng total RNA from fresh/frozen samples or 100 ng total RNA from FFPE samples. RT2 PreAMP technology uses multiplex, PCR-based preamplification to provide amplification of gene-specific cDNA target templates with minimal bias. Each RNA sample can be used to prepare enough cDNA for gene expression analysis of up to 4 different pathways.
Each RT2 lncRNA PreAMP Primer Mix is specific for one RT2 lncRNA PCR Array. During the amplification step, the RT2 lncRNA PreAMP Primer Mix enables amplification of cDNA specific for the genes targeted by the RT2 lncRNA PCR Array. A cDNA synthesis reaction from 1–100 ng total RNA from fresh/frozen samples or 100 ng–1 µg total RNA from FFPE samples provides sufficient cDNA for amplification by 4 different RT2 lncRNA PreAMP Primer Mixes, allowing gene expression analysis of up to 4 different pathways. Following preamplification, the Side Reaction Reducer eliminates residual primers. Preamplified cDNA is then ready for PCR array analysis using the appropriate RT2 lncRNA PCR Array.
Preamplification is a simple PCR reaction. Following cDNA synthesis, preamplification mix is prepared by mixing RT2 lncRNA PreAMP Primer Mix with RT2 PreAMP Mastermix, then added to cDNA template in a 96-well plate. The plate is placed in a real-time cycler, and follows a 12-cycle program detailed in the handbook. Side Reaction Reducer is added following cycling, then heat-inactivated. The amplified cDNA is stored on ice or at –20°C until use.
RT2 lncRNA PreAMP Primer Mixes are highly suited for use with small or fragmented RNA from laser-capture microdissection, fine needle aspiration biopsy, fluorescence-activated cell sorting, or FFPE samples, followed by lncRNA gene expression analysis with RT2 lncRNA PCR Arrays.
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