Additional file 2: of Germline mutation within COL2A1 associated with lethal chondrodysplasia in a polled Holstein family

Primer sequences used for validation of the mutation detected by Sanger sequencing. The heterozygous missense variant g.32476082G > R was validated using a restriction fragment length polymorphism (RFLP). Primer pairs, amplicon size (AS) in base pairs (bp), annealing temperature (AT), restriction enzyme and incubation temperature (IT) are given. Fragments in the wildtype exhibit sizes of 394 bp, 51 bp and 28 bp, whereas in the heterozygous animals, fragment sizes of 394 bp, 79 bp, 51 bp and 28 bp are present. (DOCX 14 kb)