Isolation of genomic DNA suitable for community analysis from mature trees adapted to arid environment
Language: 
English
Abstract: 

Isolation of intact and pure genomic DNA (gDNA) is essential for many molecular biology applications. It is difficult to isolate pure DNA from mature trees of hot and dry desert regions because of the accumulation of high level of polysaccharides, phenolic compounds, tannins etc. We hereby report the standardized protocol for the isolation and purification of gDNA from seven ecologically and medically important tree species of Combretaceae viz. Anogeissus (Anogeissus sericea var. nummularia, Anogeissus pendula, and Anogeissus latifolia) and Terminalia (Terminalia arjuna, Terminalia bellirica, Terminalia catappa and Terminalia chebula). This method involves (i) washing the sample twice with Triton buffer (2%) then (ii) isolation of gDNA by modified-CTAB (cetyl trimethyl ammonium bromide) method employing a high concentration (4%) of PVP (Polyvinylpyrrolidone) and 50mM ascorbic acid, and (iii) purification of this CTAB-isolated gDNA by spin-column. gDNA isolated by modified CTAB or spin-column alone were not found suitable for PCR amplification. The Triton washing step is also critical. The quality of DNA was determined by the A(260)/A(280) absorbance ratio. gDNA was also observed for its intactness by running on 0.8% agarose gel. The suitability of extracted DNA for PCR was tested by amplification with RAPD primers, which was successful. Further, rbcLa (barcoding gene) was amplified and sequenced to check the quality of extracted gDNA for its downstream applications.

Author(s): 
Gupta, Amit Kumar
Harish, null
Rai, Manoj Kumar
Phulwaria, Mahendra
Shekhawat, Narpat Singh
Item Type: 
Journal Article
Publication Title: 
Gene
Journal Abbreviation: 
Gene
Publication Date: 
11/10/2011
Publication Year: 
2011
Pages: 
156-159
Volume: 
487
Issue: 
2
ISSN: 
1879-0038
DOI: 
10.1016/j.gene.2011.06.029
Library Catalog: 
PubMed
Extra: 
PMID: 21827837

Turabian/Chicago Citation

Amit Kumar Gupta, null Harish, Manoj Kumar Rai, Mahendra Phulwaria and Narpat Singh Shekhawat. 11/10/2011. "Isolation of genomic DNA suitable for community analysis from mature trees adapted to arid environment." Gene 487: 2: 156-159. 10.1016/j.gene.2011.06.029.

Wikipedia Citation

<ref> {{Cite journal | doi = 10.1016/j.gene.2011.06.029 | issn = 1879-0038 | volume = 487 | pages = 156-159 | last = Gupta | first = Amit Kumar | coauthors = Harish, null, Rai, Manoj Kumar, Phulwaria, Mahendra, Shekhawat, Narpat Singh | title = Isolation of genomic DNA suitable for community analysis from mature trees adapted to arid environment | journal = Gene | date = 11/10/2011 | pmid = | pmc = }} </ref>