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Updated: Apr 15, 2026

Increasing cDNA Yields from Single-cell Quantities of mRNA in Standard Laboratory Reverse Transcriptase Reactions using Acoustic Microstreaming

Increasing cDNA Yields from Single-cell Quantities of mRNA in Standard Laboratory Reverse Transcriptase Reactions using Acoustic Microstreaming

Published on: July 11, 2011

Single cell transcriptome amplification with MALBAC.

Alec R Chapman¹, Zi He², Sijia Lu²

¹Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, 02138, United States of America; Graduate program in Biophysics, Harvard University, Cambridge, MA, 02138, United States of America.

|March 31, 2015

Summary

This summary is machine-generated.

We adapted Multiple Annealing and Looping-Based Amplification Cycles (MALBAC) for single-cell transcriptome amplification, achieving high detection efficiency. This method successfully analyzed the epithelial-mesenchymal transition (EMT) in mouse embryonic cells.

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Area of Science:

Molecular Biology
Developmental Biology
Genomics

Background:

Whole genome amplification of single cells is crucial for genetic analysis.
Existing methods like exponential amplification have limitations in coverage and accuracy.
Multiple Annealing and Looping-Based Amplification Cycles (MALBAC) offers quasilinear amplification for improved genome coverage.

Purpose of the Study:

To adapt the MALBAC technique for single-cell transcriptome amplification.
To evaluate the efficiency, accuracy, and reproducibility of this adapted MALBAC method.
To investigate the epithelial-mesenchymal transition (EMT) program at the single-cell level in early mouse embryos.

Main Methods:

Adaptation of Multiple Annealing and Looping-Based Amplification Cycles (MALBAC) for RNA amplification.
Single-cell RNA sequencing of cells from mouse embryos at the early gastrulation stage.
Analysis of gene expression patterns related to the epithelial-mesenchymal transition (EMT).

Main Results:

The adapted MALBAC technique demonstrated consistently high detection efficiency for single-cell transcriptomes.
The method provided accurate and reproducible transcriptome amplification and sequencing.
Successfully profiled single cells from three germ layers, including detailed examination of EMT in mesodermal cells.

Conclusions:

The adapted MALBAC technique is a powerful tool for single-cell transcriptome analysis.
This method enables high-resolution studies of developmental processes like EMT.
The findings provide new insights into cellular transitions during early embryonic development.