Twin pair 3; twin B; naive CD8 T cells
"PBMC from peripheral blood were collected from five pairs of adult MZ twins (10 total samples). The donors were all nominally healthy at sample collection, and their ages ranged from 22-27 years. The cells were first analyzed via FACS, sorted and stored at -80°C. Naïve B cells were defined by expression of CD20 and the absence of CD27. Memory B cells were defined by expression of CD20 and CD27. Naïve CD4+ T cells were defined by expression of CD4, absence of CD8 and absence of CD45RO. Naïve CD8+ T cells were defined by expression of CD8 and absence of CD4 and CD45RO. Central memory T cells were differentiate from naïve cells by expression of CD45RO and CCR7. RNA was purified from each sorted sample, and was reverse transcribed with self designed oligos. Obtained ss cDNA was purified, and the first amplification of the total transcriptome was obtained with self-designed oligos in order to add the Illumina Nextera MID P5 Adapter sequences. Enrichment for the receptor specific sequences was performed, and the Nextera P7 MID sequence was added in the final PCR."