In situ detection and genotyping of individual mRNA molecules

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Abstract
A combination of padlock probes and rolling circle amplification allows single nucleotide accuracy for transcript detection in individual cells. Increasing knowledge about the heterogeneity of mRNA expression within cell populations highlights the need to study transcripts at the level of single cells. We present a method for detection and genotyping of individual transcripts based on padlock probes and in situ target-primed rolling-circle amplification. We detect a somatic point mutation, differentiate between members of a gene family and perform multiplex detection of transcripts in human and mouse cells and tissue.