GSM2265879: SW5; Mus musculus; OTHER
Total RNA was isolated Zymo Quick RNA MicroPrep following manufacture's instructions. Two rounds of amplification were used to generate the high throughput sequencing library, the first to amplify all Igk genes using primers Igk Forward: 5’-agacgtgtgctcttccgatcGAYATTGTGMTSACMCARWCTMCA, Ck Reverse: 5’- acacgacgctcttccgatctGGATACAGTTGGTGCAGCATC.  The first round was amplified for 10 cycles, after which, this product was used to prime a 2nd round PCR with primers to add the required adapters and barcodes for the Illumina platform. Universal Forward: 5’-AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCT and Index Reverse: 5’-CAAGCAGAAGACGGCATACGAGATNNNNNNGTGACTGGAGTTCAGACGTGTGCTCTTCCGATC (where NNNNNN is a unique 6bp barcode for each sample).  This 2nd round PCR was amplified for 20 cycles.  PCR products were gel purified, mixed at equimolar ratios, and submitted to sequencing on the Illumina HiSeq (1x150bp).