Abstract

DNA was extracted using the DNeasy PowerSoil Pro kit with TBE and PBS buffers, and the results were compared with those obtained using Tris-HCl and no buffer. After extraction, DNA concentration and purity were studied using a NanoDrop spectrophotometer. The DNA extracted using PBS had a higher concentration (220.23ng/μL), while the lowest concentration was observed in TBE (45.6ng/μL). The DNA concentration measured after PCR showed that the DNA extracted using the TBE 5mL buffer had the highest concentration (708ng/μL), while the lowest concentration was in PBS (125.48ng/μL). After PCR clean-up, the final DNA concentration was higher in PBS (244.5±10.26ng/μL) and TBE 10mL (56.87ng/μL) than in Tris-HCl (38.75ng/μL), with the lowest in 5mL TBE (13.85ng/μL). The DNA sample extracted without buffer showed the lowest yield (6.18ng/μL). Statistical analysis revealed significant differences between PBS and Tris-HCl (p=7.61×10⁻⁵), with PBS-extracted DNA exhibiting a higher concentration. These results show that both buffer composition and volume are critical for yield, and that non-halogenated buffers can be effective alternatives to Tris-HCl.

Keywords:DNA extraction; Acidithiobacillus ferrooxidans; TBE buffer; PBS buffer; Tris-HCl; Quantification; Non- halogenated buffers

Abbreviations:PBS: Phosphate-Buffered Saline; TBE: Tris-Borate-EDTA; PCR: Polymerase Chain Reaction