Sequential extraction of RNA and DNA from the same sample of Neolamarckia cadamba (Roxb.) Bosser
Most of the established molecular techniques in plant molecular biology rely on the reliability of extraction methods to obtain nucleic acids from a given environmental source. The development of protocols for isolating both RNA and DNA from the same sample is recently becoming a major concern es...
| Main Authors: | , , |
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| Format: | Proceeding |
| Language: | English |
| Published: |
2007
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| Subjects: | |
| Online Access: | http://ir.unimas.my/id/eprint/10238/ http://ir.unimas.my/id/eprint/10238/1/SEQUENTIAL%20EXTRACTION.pdf |
| Summary: | Most of the established molecular techniques in plant molecular biology rely on the
reliability of extraction methods to obtain nucleic acids from a given environmental
source. The development of protocols for isolating both RNA and DNA from the
same sample is recently becoming a major concern especially when the sample is
small. Despite a number of methods that describe the simultaneous isolation of RNA
and DNA from the same sample have been reported, most of these methods have yet
to be optimized to deal with the reduced quantity and compromised quality of
samples encountered in plant genetics analysis. Neolamarckia cadamba (Roxb.) Bosser
was chosen in the present study due to its commercial value and fast growing ability.
Total RNA was successfully isolated from the leaf tissue with subsequent recovery of
DNA from the extraction mixture through alcohol precipitation. Reverse
transcriptase‐Polymerase Chain Reaction (RT‐PCR) was performed and arbitrary
primers that produced reproducible, scorable and informative bands were selected
for randomly amplified polymorphic DNA (RAPD) analysis. The amplicons of RTPCR
and RAPD were successfully obtained from all isolates, indicating that the
extracted nucleic acids were intact and pure enough for quantitative molecular
analysis. |
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