Abstract
A method for extraction of genomic DNA from sapwood tissues of mature tall trees ofPinus roxburghii, where collection of needle tissues is extremely difficult has been standardized. The extracted DNA was comparable to that obtained from the needle tissue in terms of yield and purity. The yield of extracted DNA ranged from 6.98 to 19.668 µg / 100 mg tissue and A260 / A280 ratio ranged from 1.70 to 1.87. The polymerase chain reaction (PCR) amplification of the DNA extracted from sapwood tissue using random amplification of polymorphic DNA (RAPD), inter simple sequence repeat (ISSR) and simple sequence repeat (SSR) markers was similar to that of DNA extracted from the needle tissues. Key words: Pinus roxburghii, DNA extraction, sapwood, random amplification of polymorphic DNA (RAPD), inter simple sequence repeat (ISSR), simple sequence repeat (SSR).
Highlights
Pinus roxburghii Sargent (Chir pine or long needle pine), with a long rotation of 120 years is a precious timberresin resource, native to the outer range and principal valleys of the Himalayas from Afghanistan to Arunachal Pradesh in northeastern India between 450 and 2300 m altitude
The polymerase chain reaction (PCR) amplification of the DNA extracted from sapwood tissue using random amplification of polymorphic DNA (RAPD), inter simple sequence repeat (ISSR) and simple sequence repeat (SSR) markers was similar to that of DNA extracted from the needle tissues
The cetyl-trimethylammonium bromide (CTAB) extraction buffer consisted of 100 mM Tris-HCl, 20 mM ethylenediaminetetraacetic acid (EDTA), 1.4 mM NaCl, 3% CTAB, 1% polyvinylpyrollidone (PVP) and 0.2% β-mercaptoethanol
Summary
Genomic DNA extraction from sapwood of Pinus roxburghii for polymerase chain reaction studies. A method for extraction of genomic DNA from sapwood tissues of mature tall trees of Pinus roxburghii, where collection of needle tissues is extremely difficult has been standardized. The extracted DNA was comparable to that obtained from the needle tissue in terms of yield and purity. The yield of extracted DNA ranged from 6.98 to 19.668 μg / 100 mg tissue and A260 / A280 ratio ranged from 1.70 to 1.87. The polymerase chain reaction (PCR) amplification of the DNA extracted from sapwood tissue using random amplification of polymorphic DNA (RAPD), inter simple sequence repeat (ISSR) and simple sequence repeat (SSR) markers was similar to that of DNA extracted from the needle tissues
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