AI Article Synopsis
- Barcoding in next generation sequencing is crucial for managing multiple samples and minimizing bias, but current methods face significant challenges in ChIP-sequencing.
- Transforming Y-shaped sequencing adapters into double-stranded DNA before size selection helps reduce contamination, while using qPCR to determine amplification cycles prevents over-amplification of libraries.
- The study presents a practical and affordable approach for creating barcoded ChIP-seq libraries tailored for sequencing on the Illumina platform.
Article Abstract
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.
Download full-text PDF |
Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4048252 | PMC |
| http://dx.doi.org/10.1186/1756-0500-7-312 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Want AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!