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JSIR Vol.70 [2011] >
JSIR Vol.70(11) [November 2011] >
| 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 |
| CC License: |  CC Attribution-Noncommercial-No Derivative Works 2.5 India |
| ISSN: | 0975-1084 (Online); 0022-4456 (Print) |
| Source: | JSIR Vol.70(11) [November 2011]
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