In-cell RT-PCR in a single, detached plant cell

Abstract

mRNA in situ hybridization locates specific transcripts in a tissue. PCR is ideal for detecting infrequently occurring nucleic acid sequences because of its specificity, sensitivity, simplicity, and cost effectiveness. Recently, RT-PCR in situ hybridization has been used to locate low abundance messages in a cross-sectioned tissue [6]. ln-cell PCR from mRNA has been used to detect specific transcripts in mouse and human cells [2]. The advantage of in-cell RT-PCR (reverse transcription-polymerase chain reaction) over in situ hybridization is that low abundance messages can be detected in a single detached cell. Also, because RT-PCR is performed in an intact cell, there is much less chance for detection of contaminating nucleic acid. This procedure is applicable to all single cell preparations, such as cell suspension cultures, protoplasts, and detached root border cells.