kanamycin estimation assay? range 0-50 microgram per ml - does anyone have any methods for it? thanks! (Jan/30/2008 )
Hi all..
I need a method for kanamycin estimation in ranges of 0-50 microgram/ml, does anyone have any approaches? assays? kits?
I tried a net-search.. have found a fluorescence detection method.. but not much. could anyone tel me more if they know.
thanks!
Hi,
i don't exactly understand what you want...do you want to find out how resistant your bacteria are to different concentrations of kanamycin? If so we did this: we made liquid cultures from 0 to 50 µg/ml kanamycin (every 5 µg = one culture), and put the same amout of bacteria in each tube. After 16 h we checked growth (OD600). We also did the same but with a growth curve (OD600 every 2 h) to see how fast they grow.
Is this what you want?
Stardust
i don't exactly understand what you want...do you want to find out how resistant your bacteria are to different concentrations of kanamycin? If so we did this: we made liquid cultures from 0 to 50 µg/ml kanamycin (every 5 µg = one culture), and put the same amout of bacteria in each tube. After 16 h we checked growth (OD600). We also did the same but with a growth curve (OD600 every 2 h) to see how fast they grow.
Is this what you want?
Stardust
yaa i realized wat i said cud b interpreted two ways!
i dont need what u said.. what i need is to estimate residual kanamycin in my product.. produced using bacteria grown with kanamycin. basically.. i need to estimate kanamycin in a solution.. senstive accurate assays for it. thanks!
sorry in this case i have no answer...
Well, this is not too accurate, but if you have a sensitive strain and know what its MIC is, then you can dilute the unknown and add it to a standard culture of your sensitive bacteria. With parallel controls, this could probably be made accurate within a factor of two.
people!!
i m also expecting suggestions.. like colorimetric assays, fluorescence.. Elisa.. hplc etc...!!!
thankssssssss!
people!!
i m also expecting suggestions.. like colorimetric assays, fluorescence.. Elisa.. hplc etc...!!!
thankssssssss!
"Expecting"???? What is this, some kind of test of our knowledge? (But perhaps I'm being a bit harsh.)
I was going to suggest ELISA. If you can find anti-kan monoclonals, you might be able to develop an assay. Alternately, some kind of nephelometry based on detection of kan.
people!!
i m also expecting suggestions.. like colorimetric assays, fluorescence.. Elisa.. hplc etc...!!!
thankssssssss!
"Expecting"???? What is this, some kind of test of our knowledge? (But perhaps I'm being a bit harsh.)
I was going to suggest ELISA. If you can find anti-kan monoclonals, you might be able to develop an assay. Alternately, some kind of nephelometry based on detection of kan.
OK, you can use LC/MS to qualitatively and quantitatively determine the amount of kanamycin. You'll have to be aware that there are several forms of the molecule in most commercial preparations, and know the modifications likely to be found in culture media. Oh, and you'll have to find someone with the instrument and columns and experience and interest to help you. Or you could do the MIC assay, which will probably be more relevant to what you really care about -- its activity as an antimicrobial agent.