Ecl direct labelling and detection system kit

Approximately 1 μg of gDNA was digested with HindIII restriction enzyme (Thermo Scientific) and resolved on a 0.8% agarose gel for 1 hour at 80 V. Genomic DNA fragments were transferred from the gel to a Hybond-N + membrane (Amersham) using alkaline capillary transfer. The membrane was blocked for 1 hour at 42 °C in ECL Gold Hybridisation Buffer (Amersham) supplemented with 0.5 M of NaCl and 5% (w/v) of single strand salmon sperm DNA. Hybridisation was performed at 42 °C overnight in ECL Gold Hybridisation Buffer, hybridisation probe was labelled by ECL Direct Labelling and Detection System kit (Amersham). Washing, labelling and detection were accomplished according to the manufacturer’s directions using the ECL Direct Labelling and Detection System kit (Amersham). The probe was produced via PCR of plasmid DNA pNG-Tn-pyrF using primers pyrF-1_F and pyrF-2_R (Supplementary Table S3).