Comprehensive evaluation and optimization of amplicon library preparation methods for high-throughput antibody sequencing.

ABSTRACT: High-throughput sequencing (HTS) of antibody repertoire libraries has become a powerful tool in the field of systems immunology. However, numerous sources of bias in HTS workflows may affect the obtained antibody repertoire data. A crucial step in antibody library preparation is the addition of short platform-specific nucleotide adapter sequences. As of yet, the impact of the method of adapter addition on experimental library preparation and the resulting antibody repertoire HTS datasets has not been thoroughly investigated. Therefore, we compared three standard library preparation methods by performing Illumina HTS on antibody variable heavy genes from murine antibody-secreting cells. Clonal overlap and rank statistics demonstrated that the investigated methods produced equivalent HTS dat

SUBMITTER: Menzel U 

PROVIDER: S-EPMC4014543 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature