Optimised linear amplification method for whole transcriptome analysis from low quantity RNA isolated from archival FFPE tissue
The study described a comprehensive high-fidelity method to successfully amplify low starting amounts of RNA extracted from FFPE tissue obtained from bronchial biopsies. Both amplified and unamplified RNA resulted in comparable gene expression for lung cancer and asthma samples. Overall design: FFPE samples from bronchial biopsies collected from lung cancer, asthma and asthmatic lung cancer patients were processed for RNA extraction using Recover all kit. Small amounts of RNA were amplified using in house developed random nanomer-based T3 and T7 amplification described in the study. Both amplified and unamplified RNA were subjected to whole transcriptome analysis using Ion Ampliseq human gene expression panel (Thermo Fisher Scientific, USA) and Ion S5XL sequencer.