Linseed mucilage - optimized extraction procedure for genomic research and nutraceutical applications
Keywords:
Linum usitatissimum L., mucilaginous substances extraction, DNA extraction, DNA amplification, miRNAsAbstract
Since the mucilage is released by specialized cells on the surface of the seed coat of the flax seed, some of the DNA from these cells is transferred into the mucilaginous mass. The isolation of DNA from the mucilage is greatly influenced by the conditions of mucilage extraction as well as the conditions of DNA extraction. When extracting the mucilage, it is necessary to separate the mass from the seeds by centrifugation, as pressing in gauze is likely to cause mechanical damage to the cell molecules. The temperature of the mucilage extraction is also an important factor, 40 °C was ideal for our studies, as higher temperatures release additional metabolites into the mucilage and thus make it more challenging to isolate DNA. For the actual extraction of DNA from the mucilage, a low temperature is essential as the mucilage has a higher solubility with increasing temperature. Thus, the lower the temperature, the easier it is to separate the mucilage polysaccharides from the nucleic acids. DNA purification is key for the amplification efficiency of the DNA isolated from the mucilaginous mass of flax seeds. The initial consistency of the mucilage before the isolation itself does not affect this result, although it should be noted that it acquires a consistency similar to that after the lyophilization process even by the action of liquid nitrogen on the extracted mucilage. We demonstrated the amplification efficiency of the extracted DNA using the random primers (RAPD) and sequence-specific (microRNAs).
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