title: RNA-Seq of alpha-gliadin amplicons from some Aegilops species carrying the D genome
design: For each genotype, three biological RNA samples were pooled, and cDNA was synthesized from 500 ng RNA from each pooled sample. Alpha-gliadins transcripts/amplicons were amplified from cDNA samples in two rounds of PCR reactions. The first PCR was performed using the forward and reverse alpha-gliadin specific primers including AlphaF (5-atgaaracmtttcycatc-3; for the MKTF[LP]I- motif) and AlphaR (5- ctgctgctgtgaaattrgwt-3; for the PISQQQ-motif), respectively. For each genotype, the second round of PCR was performed in 10 l reactions using fusion primers. The fusion primers included sequences necessary for Illumina sequencing. The forward fusion primer contained an 8 bp identification sequence (barcode) unique to each genotype to facilitate multiplexing and subsequent separation of sequencing reads, along with a gene-specific region. The PCR products from all genotypes were pooled, and 100 l of this pooled PCR product was sequenced using paired-end Illumina amplicon sequencing (Novogene (UK) Company).
library_name: Aegilops alpha-gliadin amplicons
identifiers.namespace: SUB15992504
identifiers.id: Aegilops alpha-gliadin amplicons