Quantitative assessment of the robustness of next-generation sequencing of antibody variable gene repertoires from immunized mice.

ABSTRACT:

Background

Next-generation sequencing (NGS) of antibody variable regions has emerged as a powerful tool in systems immunology by providing quantitative molecular information on polyclonal humoral immune responses. Reproducible and robust information on antibody repertoires is valuable for basic and applied immunology studies: thus, it is essential to establish the reliability of antibody NGS data.

Results

We isolated RNA from antibody-secreting cells (ASCs) from either 1 mouse or a pool of 9 immunized mice in order to simulate both normal and high diversity populations. Next, we prepared three technical replicates of antibody libraries by RT-PCR from each diversity scenario, which were sequenced using the Illumina MiSeq platform resulting in >106 250 bp paired-end reads per

SUBMITTER: Greiff V 

PROVIDER: S-EPMC4233042 | biostudies-literature | 2014 Oct

REPOSITORIES: biostudies-literature