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Background

RNA sequencing (RNA-seq) is a powerful method for comprehensive RNA detection, but the complexity of library preparation limits high-throughput sample processing. To overcome this, sample-specific indexed primers have been introduced during random-primed reverse transcription, allowing multiple samples to be processed in parallel (Fig.1). However, Kyoto University researchers found that RNA detection profiles depend on the index sequence used, making this approach unsuitable for accurate RNA analysis (Fig. 2, left).

Description and Advantages

The research group found that reverse transcription using random primers with specific structural features significantly reduces RNA detection bias caused by primer index sequences (Fig.2, right). This enables high-throughput analysis of multiple samples without compromising RNA detection accuracy, while substantially reducing library preparation time and reagent costs.