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Amplifying single-cell cDNA without bias

Nature Reviews Genetics volume 14, page 825 (2013)Cite this article

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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.

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References

  1. 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)

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Authors
  1. Hannah Stower
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Stower, H. Amplifying single-cell cDNA without bias. Nat Rev Genet 14, 825 (2013). https://doi.org/10.1038/nrg3631

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