PBSa spleen 20
5 prime RACE sequencing of IgH repertoire from the spleen of rainbow trout. 1 microgram of total RNA from the spleen was reverse transcribed using the SMARTER 5 RACE Kit and a CMu2 primer. Second strand synthesis was carried out to attach an illumine adaptor and purified. The cDNA library was further amplified in a two-stage PCR to produce a cDNA library with barcodes on both the 5 and 3 ends. The cDNA library was purified after each step.