Abstract
Recently, Multiple Annealing and Looping-Based Amplification Cycles (MALBAC) has been developed for whole genome amplification of an individual cell, relying on quasilinear instead of exponential amplification to achieve high coverage. Here we adapt MALBAC for single-cell transcriptome amplification, which gives consistently high detection efficiency, accuracy and reproducibility. With this newly developed technique, we successfully amplified and sequenced single cells from 3 germ layers from mouse embryos in the early gastrulation stage, and examined the epithelial-mesenchymal transition (EMT) program among cells in the mesoderm layer on a single-cell level.
Highlights
MRNA expression analyses have been extensively used in biomedical research by fluorescence in situ hybridization (FISH), qRT-PCR, and microarray, and recently have been carried out on the entire transcriptome with the advent of next-generation sequencing via RNA-seq[1]
Taking advantage of its effectiveness in DNA amplification, here we present a single-cell transcriptome amplification method based on Multiple Annealing and Looping-Based Amplification Cycles (MALBAC), named MALBAC-RNA
We developed a new single-cell transcriptome amplification method based on MALBAC
Summary
MRNA expression analyses have been extensively used in biomedical research by fluorescence in situ hybridization (FISH), qRT-PCR, and microarray, and recently have been carried out on the entire transcriptome with the advent of next-generation sequencing via RNA-seq[1]. RNA-seq has surpassed microarrays in both accuracy and dynamic range [10,11]. Gene expression is intrinsically stochastic and cannot be synchronized among cells, which leads to cell-to-cell variations in mRNA expression levels[2,4,12]. This necessitates single cell transcriptome measurements, which have prompted intense recent efforts
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