A rapid and cost-effective method of genomic DNA isolation from cyanobacterial culture, mat and soil suitable for genomic fingerprinting and community analysis

Extraction of deoxyribose nucleic acid (DNA) from plants is preliminary step in molecular biology. Fast and cost effective genomic DNA isolation from Glycine species for downstream application is a major bottleneck. Here we report a high throughput and trouble free method for genomic DNA extraction from leaf and seeds of Glycine species with high quality and quantity. Protocol reports the optimization by employing different concentrations of CTAB and PVP in extraction buffer. Efficiency of optimized protocol was compared with frequently used DNA extraction methods. Wide adoptability and utility of this protocol was confirmed by DNA extraction from leaves as well as seeds of G. max, G. soja, G. tomentella and G. latifolia. Extracted DNA was successfully subjected to PCR amplification of five microsatellite markers and four putative glycosyltransferase genes. DNA extraction protocol is reproducible, trouble free, rapid and can be adopted for plant molecular biology applications.

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Comparison of ORAGENE® and Mouthwashed-Based Saliva Collection Methods for Genomic DNA Isolation

Journal of Student Science and Technology ◽

10.13034/jsst.v8i2.77 ◽

2015 ◽

Vol 8 (2) ◽

Author(s):

Ayesha A. Hassan

Keyword(s):

Genomic Dna ◽

Dna Isolation ◽

Cost Effective ◽

Non Invasive ◽

Genomic Dna Isolation ◽

Invasive Method ◽

Saliva Collection ◽

The Difference ◽

Collection Methods

In recent years, saliva has been used as a non-invasive method of obtaining genomic DNA. Two common collection methods include mouthwash and commercially produced saliva kits. Here, a novel comparison between these two collection methods, using Scope® mouthwash and the Oragene®-Discover kit (OGR-250) from DNA Genotek Inc., was conducted to analyze differences in the quantity and quality of the DNA isolated, and cost effectiveness. The Oragene® kit yielded greater quantity of DNA, while Scope® mouthwash was more cost effective. The difference in yield was attributed to the larger volume of saliva obtained from the Oragene® kit. Isolation from both collection methods resulted in similar DNA quality. Depuis quelques années, la salive est utilisée comme une méthode non-invasive pour obtenir de l’ADN génomique. Deux méthodes de collection communes sont par rince-bouche et par des trousses commerciales de collection de salive. Ici, une comparaison entre ces deux méthodes, utilisant la rince-bouche Scope et la trousse Oragene-Discover (OGR-250) de DNA Genotek Ink, a été conduite afin d’analyser les différences dans la quantité et la qualité d’ADN isolée ainsi que dans l’efficacité du coût. La trousse Oragene a recueilli plus d’ADN, alors que Scope était moins cher. La différence en quantité est attribuée au plus grand volume de salive qui est obtenu grâce à l’Oragene. L’isolation par les deux méthodes résultait en une qualité similaire d’ADN.

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