nanoCAGE

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nanoCAGE

nanoCAGE is a modified technique of CAGE that enables genome-wide promoter analysis of small-amount of samples. Utilizing a template-switching reaction and a semi-suppressive PCR, amount of sample required is significantly reduced. nanoCAGE can accurately identify promoter sites for each transcript and obtain expression profile of respective promoters in genome-wide fashion. Several articles performing transcriptome analysis with nanoCAGE in human and mouse have been published.

Applications

  • Genome-wide gene expression analysis with tiny samples
  • Prediction of promoter sites with small amount of sample

How to choose CAGE and nanoCAGE

Library Advantage Disadvantage
CAGE Quantitative expression profile without PCR bias. More cap-dependent compared to nanoCAGE. 3μg of total RNA is required (If possible, please provide 12μg of total RNA for PCR free CAGE library preparation service).
nanoCAGE Only 50 ng of total RNA is required (100ng is recommended).
3'-end analysis with the same library is possible
PCR step during the library preparation may cause PCR biases on the expression profile.

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References

nanoCAGE

  1. C. Plessy et al, Linking promoters to functional transcripts in small samples with nanoCAGE and CAGEscan, Nat Methods, 7, 528-534 (2010)
  2. M. Salimullah et al, NanoCAGE: a high-resolution technique to discover and interrogate cell transcriptomes, Cold Spring Harb Protoc. 2011 (2011)
  3. D. Tang et al, Suppression of artifacts and barcode bias in high-throughput transcriptome analyses utilizing template switching, Nucleic Acids Res, 41, e44 (2013)
  4. M. Harbers et al, Comparison of RNA- or LNA-hybrid oligonucleotides in template-switching reactions for high-speed sequencing library preparation, BMC Genomics, 14, 665 (2013)
  5. CH. Chien, Identifying transcriptional start sites of human microRNAs based on high-throughput sequencing data, Nucleic Acids Res, 39, 9345-956 (2011)
  6. C. Plessy et al, Promoter architecture of mouse olfactory receptor genes, Genome Res, 7, 528-534 (2011)
Papers using the CAGE
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