Fig 1: Agarose gel electrophoresis showing purified 5.9 kb vector pET26b (lane V), the PCR-amplified glucose-6-phosphate dehydrogenase gene (G6PD) 1.5 kb insert (lane I), and the plasmid DNA isolated from positive transformation clones (lanes C2, C3, C4, C9, and C10), which was digested using NdeI and XhoI. Lane L is a 1 Kb Plus DNA Ladder (Thermo Fisher Scientific; catalog No. 10787018). DNA (100 ng) was quantified spectrophotometrically using Nanodrop 2000C Spectrophotometer (Thermo Scientific) and mixed with 1× DNA Gel Loading Dye (Thermo Fisher Scientific) before loading onto a 1% agarose gel (7 cm × 10 cm) and separated by electrophoresis at 120 V for 45 min in 1 × TAE buffer (40 mM Tris-acetate and 1 mM EDTA at pH 8.3) in a Mini-Sub cell GT horizontal gel electrophoresis system (Bio-Rad). The separated DNA was stained with 0.5 µg/mL of ethidium bromide, and visualized under ultraviolet light at 300–360 nm. The isolated plasmid DNA produced 2 products when digested indicating successful cloning of G6PD into the vector. The purified products were used in the subsequent ligation reaction.