Molecular Characterization and Antifungal Activity of Methanol Extract
Description
Data Supporting Molecular Characterization and Antifungal Activity of Methanol Extract of Artemisia annua leaf on Candida albicans Isolated from Three Tertiary Hospitals in Enugu State, Nigeria. Thirty azole resistant C. albicans were selected out of 500 clinical isolates for this molecular study. These were screened for detection of Candida albicans using polymerase chain reaction (PCR) technique targeting the 18S rRNA and Ergosterol II (ERGII) resistant genes. The samples were inoculated in Sabouraud dextrose broth and incubated for 48 hours. The genomic DNA was extracted from C. albicans strain using zymo research fungal/bacterial DNA mini prep kit (Zymo Research, USA) according to the manufacturer’s protocol. The master mixture for primary PCR was prepared by making a reaction mixture volume of 40 µl, made up of 1x [5 µl of PCR buffer (Sigma, Germany), 0.25 µl of Taq polymerase (Inquaba biotechnical Ind. South Africa), 1.0 µl of dNTP (100µM), 3 µl each of F1and R1, 27.75 µl of distilled water]. The master mixture in each PCR tube (40 µl) was added 10 µl of the unknown extracted DNA sample; thereby bringing the total reaction volume to 50 µl (Table 1). Each PCR tube was immediately capped after the sample was added. The used unknown extracted DNA samples and positive control sample were returned to the -20oC freezer for storage. The samples in each appropriate PCR tubes were mixed by centrifuging briefly for 7 sec. Table 2 shows the second reaction mixture of the remaining isolates with a total reaction volume of 50 µl.