Single-cell transcriptomics provides insights into the importance of stochastic transcription and facilitates more complete transcriptome characterization. Many current methods require amplification of cDNA for the analysis of low copy-number transcripts, which introduces bias. These authors optimize the amplification of cDNA libraries that are immobilized on beads. Current protocols include a step that digests primers after cDNA amplification to avoid competition with the cDNA for amplification. However, this step also degrades low copy-number cDNA, which results in amplification bias. The new protocol removes this step and washes the beads instead, which allows amplification of low copy-number transcripts at high efficiency with less bias.
References
Huang, H. et al. Non-biased and efficient global amplification of a single-cell cDNA library. Nucleic Acids Res. http://dx.doi.org/10.1093/nar/gkt965 (2013)
Rights and permissions
About this article
Cite this article
Stower, H. Amplifying single-cell cDNA without bias. Nat Rev Genet 14, 825 (2013). https://doi.org/10.1038/nrg3631
Published:
Issue Date:
DOI: https://doi.org/10.1038/nrg3631