Reference genes for quantitative Arabidopsis single molecule RNA fluorescence in situ hybridization.

ABSTRACT: Subcellular mRNA quantities and spatial distributions are fundamental for driving gene regulatory programmes. Single molecule RNA fluorescence in situ hybridization (smFISH) uses fluorescent probes to label individual mRNA molecules, thereby facilitating both localization and quantitative studies. Validated reference mRNAs function as positive controls and are required for calibration. Here we present selection criteria for the first set of Arabidopsis smFISH reference genes. Following sequence and transcript data assessments, four mRNA probe sets were selected for imaging. Transcript counts per cell, correlations with cell size, and corrected fluorescence intensities were all calculated for comparison. In addition to validating reference probe sets, we present sample preparation steps tha

SUBMITTER: Duncan S 

PROVIDER: S-EPMC10082928 | biostudies-literature | 2023 Apr

REPOSITORIES: biostudies-literature