Amplification-Free Library Preparation for Paired-End Illumina Sequencing

  • Iwanka Kozarewa
  • Daniel J. TurnerEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 733)


The library preparation step is of critical importance for the quality of next-generation sequencing data. The use of the polymerase chain reaction (PCR) as a part of the standard Illumina library preparation protocol causes an appreciable proportion of the obtained sequences to be duplicates, making the sequencing run less efficient. Also, amplification introduces biases, particularly for genomes with high or low GC content, which reduces the complexity of the resulting library. To overcome these difficulties, we developed an amplification-free library preparation. By the use of custom adapters, unamplified, ligated samples can hybridize directly to the oligonucleotides on the flowcell surface.

Key words

Next-generation sequencing Amplification-free library preparation Malaria (A  +  T)-rich genome 


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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.The Wellcome Trust Sanger InstituteCambridgeUK

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