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Title: Comparison of seven methods of DNA extraction from termitarium for functional metagenomic DNA library construction
Authors: Manjula, A
Sathyavathi, S
Gunasekaran, P
Rajendhran, J
Keywords: Cloneablity;DNA purification;Humic acid;Metagenomic DNA;Termitarium
Issue Date: Nov-2011
Publisher: NISCAIR-CSIR, India
Abstract: This study evaluated seven methods, used to isolate metagenomic DNA from termitarium, on the basis of processing time, DNA yield, purity and suitability for PCR and restriction digestion. None of the extracted DNA sample was suitable for PCR and restriction digestion. Based on higher DNA yield and lower levels of humic acids, DNA extracted using three methods were further subjected to purification. Five methods using three approaches (electro elution, silica membrane based spin column purification and gel filtration) were evaluated for purification. The simple method, employing SDS to lyse cells in situ followed by ethanol precipitation (E7) and subsequent purification using Sephadex G-50 (P5) resin yielded high concentration of cloneable DNA from termitarium sample, has been successfully used for the construction of metagenomic DNA library.
Page(s): 945-951
ISSN: 0975-1084 (Online); 0022-4456 (Print)
Appears in Collections:JSIR Vol.70(11) [November 2011]

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