Indian origin rhesus macaque MHC class-I amplicons
For each sample, we converted total cellular RNAs to cDNA using a Superscript III First-Strand Synthesis System (Invitrogen). We generated primary cDNA-PCR amplicons spanning 190 bp of exon 2 of macaque class I sequences with high-fidelity Phusion polymerase (New England Biolabs). Each PCR primer contained one of 12 distinct 10-bp MID tags along with adaptor sequences for 454 Sequencing. After purification, we normalized primary amplicons to equimolar concentrations and pooled the 48 samples into 4 groups of 12 macaques for GS FLX sequencing.
library_strategy
targeted-locus
gene
MHC class I