We have limited number of filarial worms in 70% ethanol. First I should identify the nematodes and their sex, then perform either SEM or PCR. Does clearing (lactophenol) will affect the quality of results?
In our experience in the Manter Laboratory, clearing the specimens in glycerine (glycerol) via slow evaporation of a solution of 70% ethanol and 3% glycerol (ending up with all the ethanol evaporated and only the specimen left in glycerol) provides excellent results, especially for delicate specimens. Lactophenol will destroy the DNA and should not be used for any specimens that you are planning to take DNA from later. Of course, with some filaria, you can take part of a female posterior for molecular work, and use the anterior end for morphological studies (depending on the location of the vulva and ovijector and uterus. If you use the glycerine clearing routine, the same specimen can then be cleaned, washed, and critical point dried for SEM, so you can get several layers of data from a single individual. From what we have found, glycerine does not negatively change DNA so it should be possible to extract DNA from specimens cleared in glycerine after washing with ethanol. Scott
In our experience in the Manter Laboratory, clearing the specimens in glycerine (glycerol) via slow evaporation of a solution of 70% ethanol and 3% glycerol (ending up with all the ethanol evaporated and only the specimen left in glycerol) provides excellent results, especially for delicate specimens. Lactophenol will destroy the DNA and should not be used for any specimens that you are planning to take DNA from later. Of course, with some filaria, you can take part of a female posterior for molecular work, and use the anterior end for morphological studies (depending on the location of the vulva and ovijector and uterus. If you use the glycerine clearing routine, the same specimen can then be cleaned, washed, and critical point dried for SEM, so you can get several layers of data from a single individual. From what we have found, glycerine does not negatively change DNA so it should be possible to extract DNA from specimens cleared in glycerine after washing with ethanol. Scott
Dear Scott, Thank you so much for your valuable comment. Best regards, Alireza
Dear Scott, can you tell me, please, how long each step can take approximately? I mean, how long should last the step of washing in ethanol and the step of clearing in glycerol? Thank you very much in advance. Best regards, Patricia
hi
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Article Xenomonitoring of Different Filarial Nematodes Using Single ...
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