Conservation Genetics of Conradina glabra: RADseq
Genomic DNA was converted into nextRAD genotyping-by-sequencing libraries (SNPsaurus, LLC) as in (Russello et al. 2015). Genomic DNA was first fragmented with Nextera reagent (Illumina, Inc), which also ligates short adapter sequences to the ends of the fragments. The Nextera reaction was scaled for fragmenting 30 ng of genomic DNA, although 37.5 ng of genomic DNA was used for input to compensate for the amount of degraded DNA in the samples and to increase fragment sizes. Fragmented DNA was then amplified for 27 cycles at 74 degrees, with one of the primers matching the adapter and extending 9 nucleotides into the genomic DNA with the selective sequence GTGTAGAGCC. Thus, only fragments starting with a sequence that can be hybridized by the selective sequence of the primer will be efficiently amplified. The nextRAD libraries were sequenced on a HiSeq 4000 with one lane of 150 bp reads (University of Oregon) for each 570 samples.