title: RAD-Seq of Aphis craccivora: 20 pooled whole body
design: Three g DNA from pool of 20 winged Aphis craccivora used to construct RAD library following protocol detailed in Etter & Johnson (2012). Single digestion of DNA with restriction enzyme EcoRI-HF, followed by ligation of P1 adaptors incorporating six barcodes. Barcodes were 12 nucleotides long. DNA fragments then sheared to ca. 500 bp, and run through gel electrophoresis to select samples 300-500 bp. PE-P2 adaptors then ligated to DNA fragments. Library sequenced on Illumina MiSeq, producing 6,065,758 single reads passing the filter with a read length of 250.