GBS of Pisum sativum: Accession S177 (PI381334)
The population was genotyped-by-sequencing (GBS) following the protocol proposed by Poland et al. (2012) using a combination of PstI and MspI as restriction enzymes. GBS libraries were prepared with the following modifications: a BluePippin (Sage Scientific, Beverly, MA) was used to size the libraries before PCR amplification (elution set between 50 and 65 min, on a 2% gel). Libraries were normalized, pooled, and then denatured in 0.02N NaOH and neutralized using HT1 buffer. Plate barcoding was used to enable sequencing on a shared Illumina NovaSeq S4 lane as described in Colston-Nepali et al. (2019). The pool was loaded at 225pM on an Illumina NovaSeq S4 lane using the Xp protocol according to the manufacturers recommendations. The run was performed for 2x150 cycles (paired-end mode). A phiX library was used as a control and mixed with libraries at 1% level. Base calling was performed using RTA (v3). The bcl2fastq2 software (v2.20) was then used to demultiplex samples and generate FASTQ reads.