The sequences of the TA-cloned PCR products for the validation of IS.ARG-Chip

This chip is a high-throughput quantitative PCR (HT-qPCR) platform designed to detect and quantify Insertion sequences (ISs) associated with antibiotic resistance genes (ARGs) in diverse environmental samples. A total of 114 primer sets were validated for their high specificity. The universal primers barcoding bacterial 16S rRNA gene was included as a reference gene of IS.ARG-Chip.

Specific primers were designed for each target, followed by PCR amplification. Amplicons displaying single bands of the expected fragment size were selected and subjected to T-A cloning. For each target, ten clones were randomly selected for Sanger sequencing.