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J Biomol Tech. 2014 Dec;25(4):92-5. doi: 10.7171/jbt.14-2504-001.

Protocol for optimal quality and quantity pollen DNA isolation from honey samples.

Author information

1
Departments of Botany.
2
Biotechnology, and.
3
Zoology, Mizoram University, Aizawl, Mizoram, India.

Abstract

The present study illustrates an optimized sample preparation method for an efficient DNA isolation from low quantities of honey samples. A conventional PCR-based method was validated, which potentially enables characterization of plant species from as low as 3 ml bee-honey samples. In the present study, an anionic detergent was used to lyse the hard outer pollen shell, and DTT was used for isolation of thiolated DNA, as it might facilitate protein digestion and assists in releasing the DNA into solution, as well as reduce cross-links between DNA and other biomolecules. Optimization of both the quantity of honey sample and time duration for DNA isolation was done during development of this method. With the use of this method, chloroplast DNA was successfully PCR amplified and sequenced from honey DNA samples.

KEYWORDS:

bees; chloroplast DNA; polliniferous plants

PMID:
25365793
PMCID:
PMC4211278
DOI:
10.7171/jbt.14-2504-001
[Indexed for MEDLINE]
Free PMC Article

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