Hi researchers,
I've been working with my own mucin obtained from porcine's gut and I'm trying to purify it by my own. The thing is that when I do dilutions of the fecal sample, and inoculate 100 uL in a basal medium (Na2HPO4, KH2PO4, MgCl, NaCl, NaHCO3, CaCl2, Resazurin, cysteine, NH4Cl, Vitamin and trace solution, Na2S as reducing agent) that contains 0,25% of mucin the bacteria show a growth but it's not big as expected, are like little black points after 3 days and after a week I observe the same little black points and another little points of other color (beige). The bacteria show visible growth in the 10^-3, 10^-4, 10^-5 dilutions of the sample.
I'm using the roll tubes with hungate tubes because I don't have an incubation system in the anaerobic chamber.
Nonetheless, when I do the purification of the mucin I think I'm losing too much of it so I don't know if its possible to work with the raw mucin, what do you think about it?
What could I modify of the procedure that I am doing in order to obtain positive results? and with positive results I'm talking about the possibility of observe big colonies, the main feature of this research is find bacteria that degradate the mucin.
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