Technical Considerations for Reduced Representation Bisulfite Sequencing with Multiplexed Libraries

Joint Authors

Weeks, Robert J.
Stockwell, Peter A.
Rodger, Euan J.
Morison, Ian M.
Chatterjee, Aniruddha

Source

BioMed Research International

Issue

Vol. 2012, Issue 2012 (31 Dec. 2012), pp.1-8, 8 p.

Publisher

Hindawi Publishing Corporation

Publication Date

2012-10-10

Country of Publication

Egypt

No. of Pages

8

Main Subjects

Medicine

Abstract EN

Reduced representation bisulfite sequencing (RRBS), which couples bisulfite conversion and next generation sequencing, is an innovative method that specifically enriches genomic regions with a high density of potential methylation sites and enables investigation of DNA methylation at single-nucleotide resolution.

Recent advances in the Illumina DNA sample preparation protocol and sequencing technology have vastly improved sequencing throughput capacity.

Although the new Illumina technology is now widely used, the unique challenges associated with multiplexed RRBS libraries on this platform have not been previously described.

We have made modifications to the RRBS library preparation protocol to sequence multiplexed libraries on a single flow cell lane of the Illumina HiSeq 2000.

Furthermore, our analysis incorporates a bioinformatics pipeline specifically designed to process bisulfite-converted sequencing reads and evaluate the output and quality of the sequencing data generated from the multiplexed libraries.

We obtained an average of 42 million paired-end reads per sample for each flow-cell lane, with a high unique mapping efficiency to the reference human genome.

Here we provide a roadmap of modifications, strategies, and trouble shooting approaches we implemented to optimize sequencing of multiplexed libraries on an a RRBS background.

American Psychological Association (APA)

Chatterjee, Aniruddha& Rodger, Euan J.& Stockwell, Peter A.& Weeks, Robert J.& Morison, Ian M.. 2012. Technical Considerations for Reduced Representation Bisulfite Sequencing with Multiplexed Libraries. BioMed Research International،Vol. 2012, no. 2012, pp.1-8.
https://search.emarefa.net/detail/BIM-991871

Modern Language Association (MLA)

Chatterjee, Aniruddha…[et al.]. Technical Considerations for Reduced Representation Bisulfite Sequencing with Multiplexed Libraries. BioMed Research International No. 2012 (2012), pp.1-8.
https://search.emarefa.net/detail/BIM-991871

American Medical Association (AMA)

Chatterjee, Aniruddha& Rodger, Euan J.& Stockwell, Peter A.& Weeks, Robert J.& Morison, Ian M.. Technical Considerations for Reduced Representation Bisulfite Sequencing with Multiplexed Libraries. BioMed Research International. 2012. Vol. 2012, no. 2012, pp.1-8.
https://search.emarefa.net/detail/BIM-991871

Data Type

Journal Articles

Language

English

Notes

Includes bibliographical references

Record ID

BIM-991871