Sula nebouxii_lociPCR_female_blood_13
DNA samples were standardized to 10 ng/ul. We amplified the autosomal, Z-linked and W-linked loci in the same PCR reaction using the Phusion Flash High Fidelity from Thermo Fisher Scientific (Cat. No. F548L). PCR products were purified using Agencourt AMPure XP (Cat. No. A63882). PCR products were multiplexed and sequenced in a NextSeq 500 Illumina machine (paired-end, 75 nucleotides long) at UNAM. The quality of the reads was verified using FastQC, and the remaining adaptors and low-quality bases were removed with Trimmomatic (v.036. Reads were aligned using bowtie2 (v. 2.3.4.1) against the genome sequence of the Magnificent Frigatebird or the transcriptome assemblies of the Blue-footed Booby. We used samtools to extract the reads that mapped uniquely to the expected loci.