Identification of cell wall type is an essential step for the characterization of actinomycetes. I have prepared the cell wall hydrolysate using 6N HCl and carried out the silica gel TLC with the mobile phase of Methanol-DH2O-6N HCl-pyridine (80:26:4:10) and visualized the spots using ninhydrin spray. I couldn't get the amino acids separated. Is there any problem with my TLC? If you have any protocol which is good for the detection of LL-DAP and meso-DAP means share it?
Hi Muthamil,
Prior to testing a new eluent system I would recommend to perform a 2D TLC by just turning the card around - preferably use a 20 x 20 cm card. In addition, I would reommend to try 1% of sulfuric acid in pure ethanol in combination with hot air, e.g. (hair) dryer, to make the dots visible. You can also do two cards, one for ninhydrin and one for H2SO4. The latter has the advantage that any organic compounds are turned to ashes which is visible as brown to black colors.
Regards,
Alec
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