soybean Noir1-SGC-01 Exome sequencing
Illumina Paired End libraries (Illumina) were constructed using Illumina’s PE Kit (part no. PE-102-1001). A custom microarray was built for soybean exome sequence capture based on the soybean gene annotation by Roche NimbleGen (design 100310_Gmax_public_exome_cap_HX3). The library exon sequences were captured by hybridizing to the microarray for 72 h at 42°C in the presence of 20 µL of plant capture enhancer per subarray. Slide washing and sample library elution were performed as described by Fu et al. (Plant J 62: 898-909). Posthybridization amplification consisted of 16 cycles of PCR. Following the completion of the amplification reaction, the samples were purified using a Qiagen Qiaquick column using the manufacturer’s recommended protocol. The resulting postcapture enriched sequencing libraries were diluted to 10 nm and used in cluster formation on an Illumina cBot, and paired-end sequencing was done using Illumina’s Genome Analyzer IIX. Both cluster formation and 76-bp paired-end sequencing were performed using the Illumina protocols.