Isolation and PCR amplification of genomic DNA from dried capsules of cardamon (Elettaria cardamomum M.) |
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Authors: | S Syamkumar Mrudula Jose B Sasikumar |
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Institution: | (1) Division of Crop Improvement and Biotechnology, Indian Institute of Spices Research, Calicut, 673012 Kerala, India |
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Abstract: | Cardamom is an important spice, condiment and medicine, and international commodity. DNA-based molecular profiling will be
aid in protecting the intellectual property rights of those who trade cardamom on the world market. Commercial cardamom has
so far proven recalcitrant to traditional DNA extraction methods. In this paper we report a protocol for the isolation of
amplifiable genomic DNA from traded cardamom. The method involves a modified CTAB (hexadecyltrimethylammonium bromide) extraction
step, followed by a purification step to remove polysaccharides, proteins, and polyphenols, which are abundant in storage
tissue such as cardamom capsules. The yield of DNA was 6–7 μg g−1 tissue. Spectrophotometric and electrophoretic analysis indicated that the isolated DNA was highly pure and of high molecular
weight. The isolated DNA could be amplified using different random decamer primers. The protocol has trade implications as
it will help in the PCR-based characterisation of traded cardamom. This protocol can be further extended to develop Sequence
Characterised Amplified Regions (SCAR) markers for profiling cardamoms. |
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Keywords: | Elettaria cardamomum cardamom DNA isolation RAPD PCR |
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