Journal of Applied Biosciences (J. Appl. Biosci.) [ISSN 1997 - 5902]
Volume 21: 1237 - 1245. Published September 7, 2009.
Microsatellite DNA marker isolation from Picrorhiza kurrooa Royle ex Benth by magnetic capture
Hussain, M.A.*, Aggarwal, R.K.**, Bedi, Y.S.,* and Abdin, M.Z.***
* Plant Biotechnology Division, Indian Institute of Integrative Medicine, Canal Road, 180001, Jammu Tawi, India.
**Centre for Cellular and Molecular Biology, Hyderabad, 500007, India.
***Centre for Transgenic Plant Development, Department of Biotechnology, Hamdard University, New Delhi, 110062, India.
ABSTRACT
Objective: To establish enriched libraries by magnetic capture of microsatellites.
Methodology and results: Microsatellite containing sequences were isolated from enriched genomic libraries of Picrorhiza kurrooa Royle ex Benth. The enrichment was based on magnetic capture of microsatellite motifs by streptavidin coated magnetic beads and mixed biotin labeled probes hybridization capture strategy. Out of 96 colonies randomly picked for sequencing, 152 loci containing unique SSR motifs were identified. The majority of these (70%) were dinucleotide and trinucleotide repeats. The GA/GT repeat was the most common accounting for 58% of all repeat types. The average clone size of the positive clones was 500 to 1000 bp. The enrichment efficiency obtained by magnetic capture was 75% as compared 0.4 to 0.5% realized with conventional colony hybridization protocols.
Conclusion and application: Microsatellite sequences (EU883601 to EU883622 and FJ617210 to FJ617223) were successfully, and for the first time, isolated from the endangered medicinal herb Picrorhiza kurrooa Royle ex Benth. (Royle, 1835). These markers can be applied in the assessment of genetic similarity or diversity in order to assign specimens to heterotic groups on the basis of repeat length polymorphism.
Key words: magnetic capture, microsatellites, molecular marker, Picrorhiza kurrooa, SSR
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