Tbe buffer

The PCR products for α-thalassaemia were analysed by using agarose gel electrophoresis. PCR products of multiplex gap PCR were run on a 1.5% agarose gel electrophoresis using 1× Tris-Borate-EDTA (TBE) buffer. Whereas, PCR products for α-thalassaemia from multiplex ARMS PCR were run on a 1.2% agarose gel.
The PCR products from the β globin gene, except for reaction in MARMS-A, MARMS-B and MARMS-F, were separated using 2% agarose D1, Low EEO Pronadisa (Laboratorios CONDA, Madrid, Spain) in 1X TBE buffer (Biobasic, Markham, ON, Canada). DNA band was visualized under transilluminator (Vilber Lourmat, Sud Marne-la-Vallée, France). QIAxel Advanced System (QIAGEN GmBH, Hilden, Germany) was used to analyse MARMS-A, MARMS-B, and MARMS-F which used automated capillary electrophoresis to separate the products based on their sizes and visualised on the interfaced computer system using its dedicated QIAxcel ScreenGel Software (QIAGEN GmBH, Hilden, Germany).

DNA was extracted using cell lysis method [13 ]. Briefly, 40 μL of SCLB (1 mL of 1× concentration of Tris-Borate-EDTA [TBE] buffer [Bio Basic, Canada] plus 10 μL of proteinase K [5 mg/mL]) were added to individual sterile polymerase chain reaction (PCR) tubes. A single colony of E. coli from MHA was added as DNA template. Amplification was carried out using in BioRad MyCycler™ (Bio-Rad, California, USA) at 80°C for 10 min followed by one cycle at 55°C for 10 min. Then, 80 μL of ddH₂O was added, centrifuged at 7000 rpm for 2 min, and finally, the extracted templates were stored at 4°C until used.