School Of Basic And Applied Sciences

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Author: search.filters.author.Yadav, Hemendra

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    Combing of picogram level DNA equivalent to genomic DNA present in single human cell by self propelled droplet motion over a stable gradient surface
    (Academic Press Inc., 2023-11-08T00:00:00) Yadav, Hemendra; Algaonkar, Prashant S.; Chakraborty, Sudip; Ramakrishna, Wusirika
    DNA combing is a powerful technique for studying replication profile, fork-directionality and fork velocity. At present, there is requirement of a methodology to comb DNA present in a single human cell for studying replication dynamics at early embryonic stage. In our study, a surface having dual characteristics i.e., affinity towards negatively charged single DNA molecules and a hydrophobic gradient for self propelled droplet motion of combing solution was developed. The surface was made by coating of TCOS (trichloro-octylsilane) by vapor diffusion on APTES (Aminopropyl-triethoxysilane) coated glass slides. A gradient surface having high deposition efficiency (DE) was developed on which 5 picogram DNA equivalent to genomic DNA present in one single human cell can be combed. The gradient surface was thermostable in nature having the ability to sustain boiling temperature for two hours and sustain anisotropy in 70 % ethanol for 80 h. Applicability for multiple runs was enhanced such that the surface can be used for 13�14 times. Factors associated with gradient surface are unidirectional movement of combing solution droplet over the gradient surface for combing straight DNA molecules and a longer gradient surface of more than 1 cm such that long size DNA molecules can be combed. Ellipsometry and contact angle hysteresis confirmed the presence of hydrophobic gradient. XPS (X-ray photoelectron spectroscopy) and FTIR (Fourier Transform Infrared Spectroscopy) confirmed the presence of characteristic affinity towards negatively charged DNA molecules on the gradient surface. Combing solution was optimized for increasing deposition efficiency and for increasing the applicability of gradient surface for multiple runs. High temperature of combing solution was found to increase Deposition Efficiency. Combing solution was also optimized for combing single DNA molecules over the gradient surface. Single DNA molecules were combed by reducing pH and lowering concentration of triton-X in the combing solution. Dye: bp ratio was optimized for high fluorescent intensity and low surface background. � 2023 Elsevier Inc.
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    Multi-dimensional Studies on Thermotolerant Phosphate Solubilizing Bacteria Isolated from Rock Phosphate Mines for Bioactivation of Rock Phosphate
    (Taylor and Francis Ltd., 2022-08-24T00:00:00) Yadav, Hemendra; Karkho, Richard; Mathur, Sujata
    We conducted multi-parametric studies on thermotolerant phosphate solubilizing bacteria useful in bioactivation of rock phosphate. Selected two different thermotolerant phosphate solubilizing cultures. Statistical optimization found pH, ammonium sulfate and EDTA concentration to be the significant variables for high phosphate solubilization by Bacillus sp. BISR-HY63. Alkaline phosphatase production by Brevibacillus sp. BISR-HY07 was increased from 1400 unitsL?1 to 3000 unitsL?1. Partial purification of alkaline phosphatase performed by size exclusion and anion-exchange chromatography and the enzyme purified by 76 folds. Organic compost prepared and inoculated with alkaline phosphatase producing bacterium BISR-HY07 for increasing it�s efficacy. Soluble P decreased from 9.5 mgKg?1 to 4.1 mgKg?1 and available P increased from 200 mgKg?1 to 1000 mgKg?1 after incubation of 50 days. When cultures used as bioinoculants, total P content absorbed by the plants inoculated by Brevibacillus sp. BISR-HY07 along with rock phosphate was highest and significantly higher at 9.3 gkg?1. Shoot length of plants treated by rock phosphate and Brevibacillus sp. BISR-HY07 found to be highest and significantly higher at 31.5 cm. Root length, shoot biomass and root biomass of plants treated by rock phosphate and Bacillus sp. BISR-HY63 was highest but not significantly high at 10.3 cm, 0.73 g and 0.34 g respectively. � 2022 Informa UK Limited, trading as Taylor & Francis Group.