title: Protocol for optimal quality and quantity pollen DNA isolation from honey samples.
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.
authors.affiliation: Departments of Botany.
authors.forename: Ralte
authors.initials: R
authors.lastname: Lalhmangaihi
authors.affiliation: Biotechnology, and.
authors.forename: Souvik
authors.initials: S
authors.lastname: Ghatak
authors.affiliation: Departments of Botany.
authors.forename: Ramachandra
authors.initials: R
authors.lastname: Laha
authors.affiliation: Zoology, Mizoram University, Aizawl, Mizoram, India.
authors.forename: Guruswami
authors.initials: G
authors.lastname: Gurusubramanian
authors.affiliation: Biotechnology, and.
authors.forename: Nachimuthu Senthil
authors.initials: NS
authors.lastname: Kumar
mesh_terms: D000818:Animals; D001516:Bees / Q000502:physiology; D002736:Chloroplasts / Q000235:genetics; D018742:DNA, Chloroplast / Q000302:isolation & purification*; D005819:Genetic Markers; D006722:Honey / Q000032:analysis*; D011058:Pollen / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA
chemical_list: D018742:DNA, Chloroplast; D005819:Genetic Markers
keywords: bees; chloroplast DNA; polliniferous plants