RT-PCR analysis of three active genes located in the neocentromeres. (Lane 1) Primer 311-92 amplified an 840-bp fragment from gene G311 in Seneca 60 (maize), Sun II (oat), OMA3.01 line, and neoM3 line. This fragment spans exon 1 to exon 5 of G311. (Lane 2) Primer G311-15 amplified a 707-bp fragment from gene G311 in all four lines. This fragment spans exon 2 to exon 7 of G311. (Lanes 3,4,5,6) Four different maize-specific primers were designed to amplify different parts of G311 in the two oat-maize chromosomal addition lines. (Lanes 7,8) Two different maize-specific primers were designed to amplify gene G576 in the two oat-maize chromosomal addition lines. Note: Both primers amplified two bands in Seneca 60, suggesting that G576 has a homologous gene of G576 in a different maize chromosome. Both primers, however, amplified a single band in the two addition lines. (Lanes 9,10) Two different maize-specific primers were designed to amplify gene G959 in two oat-maize chromosomal addition lines. All primer sequences are provided in Supplemental Table S3.
