title: Degenerative primer design and gene sequencing validation for select turkey genes.
abstract: We successfully designed and validated degenerative primers for turkey genes MUC2, RPS13, TBP and TFF2 based on chicken sequences in order to use gene transcription analysis to evaluate (quantify) the mucin transcription to probiotic supplementation in turkeys. Primers were designed for the genes MUC2, TFF2, RPS13 and TBP using a degenerative primer design method based on the available Gallus gallus sequences. All primer sets, which produced a single PCR amplicon of the expected sizes, were cloned into the TOPO(®) vector and then transformed into TOP 10(®) competent cells. Plasmid DNA isolation was performed on the TOP10(®) cell culture and sent for sequencing. Sequences were analyzed using NCBI BLAST. All genes sequenced had over 90% homology with both the chicken and predicted turkey sequences. The sequences were used to design new 100% homologous primer sets for the genes of interest.
authors.affiliation: The Ohio State University, Department of Animal Science.
authors.forename: Stephanie L
authors.initials: SL
authors.lastname: Hutsko
authors.affiliation: The Ohio State University, Department of Animal Science lilburn.1@osu.edu.
authors.forename: Michael S
authors.initials: MS
authors.lastname: Lilburn
authors.affiliation: The Ohio State University, Department of Animal Science.
authors.forename: Macdonald
authors.initials: M
authors.lastname: Wick
mesh_terms: D000818:Animals; D030161:Avian Proteins / Q000235:genetics* / Q000378:metabolism; D017931:DNA Primers / Q000235:genetics*; D016133:Polymerase Chain Reaction; D014422:Turkeys / Q000235:genetics
chemical_list: D030161:Avian Proteins; D017931:DNA Primers
keywords: cloning; homology; primers; sequencing; transcription