A method for generating highly multiplexed ChIP-seq libraries

BMC Research Notes

Dimitris Thanos¹  Antonis Kokkalis¹  Chrysa Nikopoulou¹  Ethan Ford¹ 

[1] Biomedical Research Foundation, Academy of Athens, 4 Soranou Efesiou Street, Athens 11527, Greece

关键词: Epigenetics;    Genomics;    Next generation sequencing;    ChIP-seq;    Chromatin immunoprecipitation;   

Others  :  1132700
DOI  :  10.1186/1756-0500-7-312

 received in 2014-02-17, accepted in 2014-05-07,  发布年份 2014

【 摘 要 】

Background

The barcoding of next generation sequencing libraries has become an essential part of the experimental design. Barcoding not only allows the sequencing of more than one sample per lane, but also reduces technical bias. However, current barcoding strategies impose significant limitations and/or technical barriers in their implementation for ChIP-sequencing.

Findings

Converting Y-shaped sequencing adapters to double stranded DNA prior to agarose gel size selection reduces adapter dimer contamination and quantitating the number of cycles required for amplification of the library with qPCR prior to library amplification eliminates library over-amplification.

Conclusions

We describe an efficient and cost effective method for making barcoded ChIP-seq libraries for sequencing on the Illumina platform.

【 授权许可】

   
2014 Ford et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

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