[Leish-l] S.O.S.

[tiago.rf at usp.br]

Dear Rita, 
It seems like any of the suggested protocols may work, because we have done several transfections of L. braziliensis using the 450V 500uF protocol in the lab (Kapler et al 1990) , using strains MHOM/BR/75/M2904 and MHOM/BR/94/H3227. It depends really on what you are doing and which efficiency it requires. The Amaxa T-cell protocol has the best efficiency, but you will need their Nucleofector system. 
Have you checked the IC50 of G418 for your specific strain? We have found it is dependent on strain and batch used. 

All the best, 

Tiago Rodrigues Ferreira 
Laboratório de Parasitologia Molecular 
Faculdade de Medicina de Ribeirão Preto 
Universidade de São Paulo 
Av. Bandeirantes, 3900 - CEP 14049-900 
Ribeirão Preto - SP - Brasil 
+55 (16) 3602 3053 

----- Mensagem original -----

> De: "Rita Mukhopadhyay" <rmukhop at fiu.edu>
> Para: "Patrick Bastien" <patrick.bastien at univ-montp1.fr>,
> leish-l at lineu.icb.usp.br
> Enviadas: Terça-feira, 18 de Fevereiro de 2014 3:40:44
> Assunto: Re: [Leish-l] S.O.S.

> Dear Colleagues,

> Can anyone please send me transfection protocols for braziliensis and
> panamensis? We are trying with G418 selection. Looks like they are
> too sensitive to the regular 1500V electroporation protocol.

> Thanks a lot for your help.

> Rita Mukhopadhyay
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