Abstract
Isolation of high quality RNA from ripening banana fruit tissue is difficult due to high levels of polysaccharides and other substances that interfere when using conventional procedures for RNA isolation. These substances not only decrease the yield but the quality of RNA is almost unusable. We describe here a simple RNA procedure that effectively removes these contaminating substances without affecting the yield. Following this procedure, we routinely obtained 80–150 μg of total RNA per g fresh tissue. The RNA is of good quality and suitable for northern analysis, RT-PCR and cDNA library construction.
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Abbreviations
- AMVRT:
-
Avian Myelosblastosis Virus Reverse Transcriptase
- CIP:
-
Calf Intestine Alkaline Phosphatase
- CTAB:
-
cetyltrimethylammoniumbromide
- DEAE:
-
diethylaminoethyl
- DEPC:
-
diethyl pyrocarbonate
- EtBr:
-
ethidium bromide
- EtOH:
-
ethanol
- RT-PCR:
-
reverse transcription polymerase chain reaction
- SDS:
-
sodium dodecyl sulphate
- SSC:
-
saline sodium citrate
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NBRI publication No. 488(NS).
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Asif, M.H., Dhawan, P. & Nath, P. A simple procedure for the isolation of high quality RNA from ripening banana fruit. Plant Mol Biol Rep 18, 109–115 (2000). https://doi.org/10.1007/BF02824018
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DOI: https://doi.org/10.1007/BF02824018