Electrophoresis was performed in TAE buffer with a constant voltage of 200 V at 60°C for 240 min. The DNA bands were stained by SYBR green I (Amresco, Solon, OH) and were photographed with a UVI gel documentation system (UVItec, Cambridge, UK).
Electrophoresis was performed in a 1 × TAE buffer with a constant voltage of 80 V for 17 h at 60°C using a DCode Mutation Detection System (Bio-Rad, USA).
The reaction mixture was separated on a 2% agarose gel prepared in gel buffer (20 mM Na2HPO4; pH 7.0) with a voltage of 14 Vcm-1 applied for 30 min. For Cy3 detection, the gel was scanned with the Typhoon 8600 Variable Mode Imager (Amersham Biosciences Europe, Freiburg, Germany).
The enzyme activity was verified in a 1 % agarose gel in TAE buffer with ethidium bromide (0.5 μL/mL), with a voltage of 1 2.5 volt/cm.
PFGE gels were run in 0.5% Tris borate EDTA buffer at 12°C with an angle of 120° with a voltage of 3 V/cm and switch times of 300 s for 120 h.
Take half of the sample from Step 63 (20 μl) and run the DNA digest using gel electrophoresis with a 1.0% 1xTBE agarose gel, 1xTBE buffer, and a constant voltage setting of 6V/cm for approximately 2 hours.
The accelerating voltage was 20 kV, with a grid voltage of 85.0%, guide wire voltage of 0.050%, and a delay of 100 ns.
The accelerating voltage was 19 kV with a grid voltage of 75.2%; the mirror voltage ratio was 1.12, and the acquisition mass range was 200 3,000 Da.
IEF was carried out with a starting voltage at a maximal voltage of 500 V for various time intervals, and the IPG strips were desalted by soaking the IPG strips in rehydration buffer or distilled water twice for 5 min, a procedure herein named in-gel dialysis.
