Celso,
in our experience, the wet-weight of 10 worms is about 0.2g (mixed sex).
In the past, we developed a Direct-PCR protocol using total worms or even
total larval stages without prior DNA extraction. In the meantime this
protocol has been used by Helene Monet from Perpignan (France) and Martin
Kalbe from Erlangen (Germany) successfully. However, we have not yet tried
to use this protocol for RT-PCR which might be difficult. For RT-PCR, you
should consider to extract the RNA from individual worms using common kits
like RNAeasy from Qiagen or the Dynabead magnetic-particle procedure
(Dynal). Both work well in our hands.
Here are some references which might be helpful in this respect:
1: Grevelding, C.G., Kampkoetter, A. and Kunz, W. (1997) Schistosoma
mansoni: PCR on adult and larval stages without DNA extraction.
Experimental Parasitology 85, 99-100
2: Grevelding, C.G., Kampkoetter, A., Hollmann, M., Schaefer, U. and Kunz,
W. (1996) Direct PCR on fruitflies and blood flukes without prior DNA
isolation. Nucleic Acids Research 24, 4100-4101
3: Schuessler, P., Gohr, L.G., Sommer, S., Kunz, W. and Grevelding, C.G.
(1995) A simple method for the combined isolation of nucleic acids and
protein from minor amounts of tissue. Trends in Genetics 11, 378-379
Sincerely,
Christoph G. Grevelding
>Hello all,
>>Please, I need data about weight of schistosoma mansoni worm total weight
>(average for males and females). Exist such information?
>>In another hand, I looking information about PCR and RT-PCR protocols for
>individual schistosoma woms. If you have any experience in this field,
>please help me.
>>Many Thanks
>>Celso Romero
