Description
Analysis of small amounts of RNA is critical for biopsy, FFPE, and other types of samples. RT 2 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. RT 2 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 RT 2 lncRNA PreAMP Primer Mix is specific for one RT 2 lncRNA PCR Array. During the amplification step, the RT 2 lncRNA PreAMP Primer Mix enables amplification of cDNA specific for the genes targeted by the RT 2 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 RT 2 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 RT 2 lncRNA PCR Array.
Preamplification is a simple PCR reaction. Following cDNA synthesis, preamplification mix is prepared by mixing RT 2 lncRNA PreAMP Primer Mix with RT 2 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.
