Methanol fixation and tagmentation of RNA/DNA hybrids directly enable single-cell transcriptome sequencing

Methanol fixation enables preservation of RNA and facilitates <i>in situ</i> reverse transcription of full-length cDNA. Importantly, cells maintain their integrality after reverse transcription and transposition even under low concentration of methanol. Reducing the methanol concentration to 40% further enhances transcript capture efficiency. At the single-cell level, this strategy enables the capture of full-length transcriptomes, demonstrating a great potential for application in single-cell sequencing.