Ig-seq Mus musculus CD138-3T3-C1-wt1
Total RNA was purified from splenocytes of the mice using RNeasy plus mini kit (Qiagen) according to the manufacturers protocol. The 5 RACE approach was used for the generation of cDNA library to eliminate errors caused by multiplex primers. cDNA synthesis products were treated with uracil-DNA glycosylase (New England BioLabs, 5 U per reaction). Two-stage PCR amplification was carried out to amplify the IgG-specific antibody variable region. Illumina adaptors were ligated using Ovation Ultralow Library System V2 (NuGEN) according to the manufacturers protocol. The DNA concentration of the library was measured using Qubit and Real-time PCR, and libraries were submitted for a final quality control step on a Bioanalyzer 2100 (Agilent) prior to sequencing.