Improved tagmentation-based whole-genome bisulfite sequencing for input DNA from less than 100 mammalian cells

Abstract

Aim: To develop a whole-genome methylation sequencing method that fulfills the needs for studies using ultra-low-input DNA. Materials & methods: The tagmentation-based whole-genome bisulfite sequencing (T-WGBS) technology is modified, enabling stable library construction with complexity from minimally 0.5 ng of initial genomic DNA, which equals less than 100 mammalian cells. Results: We thoroughly assessed the performance of this T-WGBS method by sequencing the methylomes of a rice strain and pre-implantation embryos of rhesus monkey and compare to traditional WGBS approach, thereby demonstrating the efficacy of this new approach. Conclusion: This new approach is highly attractive for the complete methylome analysis of very few cells, for example, mammalian pre-implantation embryos, or tiny human biopsy specimens.

Keywords::

• DNA methylation
• T-WGBS
• Tn5 transposase
• WGBS

Acknowledgements

The authors thank Andrei Alexeev for helpful discussion on the design of this method.

Financial & competing interests disclosure

This project was funded by the Guangdong Natural Science Foundation (No. S2013010016525), the National High Technology Research and Development Program of China (No. 2012AA02A201) and Shenzhen Key Laboratory of Transomics Biotechnologies (No. CXB201108250096A). The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

No writing assistance was utilized in the production of this manuscript.

Ethical conduct of research

The authors state that they have obtained appropriate institutional review board approval or have followed the principles outlined in the Declaration of Helsinki for all human or animal experimental investigations. In addition, for investigations involving human subjects, informed consent has been obtained from the participants involved.

Additional information

Funding

This project was funded by the Guangdong Natural Science Foundation (No. S2013010016525), the National High Technology Research and Development Program of China (No. 2012AA02A201) and Shenzhen Key Laboratory of Transomics Biotechnologies (No. CXB201108250096A). The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed. No writing assistance was utilized in the production of this manuscript.