title: C_americana
design: I used a double digest restriction site associated DNA sequencing (ddRADseq) protocol to obtain SNPs from across the genome (Peterson et al. 2012). I performed DNA extractions on either fresh or dried tissue in 96-well plates using a modified CTAB chloroform extraction protocol. Next gen sequencing libraries were built with a Biomek NXp liquid handling robot. Samples were randomized within plates, then digested with the enzymes of ApoI HF and Sph! HF simultaneously. I then ligated one of 48 unique adapters to the DNA of each sample, purified with ampure beads, and size selected for fragments of 450-550bp using a BluePippin machine. After size selection, I made 9 pools of 48 samples, each with unique adapters, and performed PCR on each pool using a different primer for each one, given each sample a unique combination of indices. PCR products were normalized based on qPCR, then pooled and sent to BGI for sequencing on two lanes of Illumina HiSeq 4000. Samples here are demultiplexed.
library_name: PA95_1