I am working on the Mucilagenous fresh water algae (Desmid) and the protocol followed was using CTAB buffer without using Liquid nitrogen. When pellet is dissolved in pure water, after sometime it formed a thick gel like material, pipetting was not even possible, any how during gel electrophorosis process, there are some DNA.
What could be the possible solution to remove the mucilage from my finished product or during extraction process.
Dear Ahmed Sir,
Thank you for taking your valuable time for answering my query. In this case I think the mucilage is produced by algae itself and is heavily embedded in it, which even during centrifugation is hard to remove. During DNA extraction process I have incubated the homogenized sample in CTAB and Centrifuged at 12000 rpm for 10 min to pellet the debris then added equal volume of Chloroform: Iso amyl alcohol (24:1 v/v) to the supernatant and gently vortexd for 10 min and centrifuge 13000 rpm for 10 min.
After precipitation and washing etc quantification of DNA is checked by Nanodrop spe
ctrophotometer with absorbance ratio OD [260]/ OD [280] which gave the value between 1.826 and 1.930, concentration of dsDNA is between 60-81 (ng/ul).
Is it possible to remove the gelatineous material from DNA, kindly help.
Dear Sir,
Thank you once again, as with regard to your query, yes the gelatinous material is soluble in 70% alcohol. No I haven't tried phenol or trizol before, but i will try to purify the sample using DNA kit as suggested by you, I will let you know the outcome of it as early as possible.
Thank you
Hi,
"Mucilagenous" is because Polysaccharides. I had this problem before on Corchorus oltorius plants. I did that: (1) cultivated seeds in the whole dark until emerging seedlings to reduce phenols and poly saccharides; (2) repeated Chloroform: Isoamyl alcohol (24:1 v/v) several time in DNA Extraction.
It is just.
Potassium acetate precipitations work nicely to precipitate out polysaccharides. Just remember your DNA/RNA is in solution! Follow with an ethanol precipitation.
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