ABSTRACT
Aims: Isolation and biochemical characterization of yeasts from toddy and standardization of best method for DNA extraction from yeast. Study Design: Biochemical characterization of yeast and genomic DNA extraction by manual and kits methods. Place and Duration of Study: Department of Microbiology, Institute of Genetic Engineering, Badu, kol-128, India and Molecular Mycopathology Lab, P. G. Department of Botany, Ramakrishna Mission Vivekananda Centenary College, Rahara, kol-118, India, from November 2012 –April,2013. Methodology: Toddy was collected in sterilized polythene bags from palm tree (Borassus flabellifer L; Family: Arecacea) in the morning, from Badu, 24-parganas (N) India. Isolation of yeasts was done by the method of Beech and Davenport [15] using MA (Malt extract) medium. Biochemical Identification was performed by using basal medium and procedure [1,2,15]. Genomic DNA extraction was done by manual and kits methods (UniflexTM DNA isolation Kit). Quality of extracted DNA was checked by the absorbance ratio (A260 / A280) ranged from 1.8 to 2.0. Results: By performing morphological, microscopical and biochemical characterization the isolated yeast from toddy was identified as Candida famata consulting with the key of yeast published [1,2]. The UniflexTM DNA isolation Kit method is much more convenient way to get pure and high quality DNA than the manual methods. Conclusion: Isolated yeast from toddy was identified as Candida famata. The genomic DNA of Candida famata was extracted purely by UniflexTM DNA isolation Kit. This method was better and more convenient than manual method.
ABSTRACT
OBJECTIVE: To develop a kind of Agkistrodon acutus (Günther) DNA test kit, evaluate its quality indexes including specificity, stability, sensitivity and repeatability, and inspect the qualities of commercialAgkistrodon acutus (Günther) samples. METHODS: The Agkistrodon acutus (Günther) DNA test kit was developed and modified according to the method recorded in Chinese Pharmacopoeia (2010 edition). EighteenAgkistrodon acutus (Günther) samples were randomly collected from Beijing, Tianjin, and Changchun. The kit assay was performed to identify these samples with the pharmacopoeia method as the standard control. RESULTS: The kit proved effective after 20 times of freezing and thawing, and repeatability test showed same data for three tests. The lower limit of quantitation was 0.025 g. The specificity test confirmed that 14 samples were genuine, and 4 were adulterants. All of the identification results by the kit assay were in accordance with the ones by the pharmacopoeia method. CONCLUSION: The developed DNA test kit is accurate and effective for identification of Agkistrodon acutus (Günther). Compared with the pharmacopoeia method, it is simpler and more rapid, demonstrating broad prospect in quality inspection of Agkistrodon acutus (Günther).