<html><head><style type='text/css'>p { margin: 0; }</style></head><body><div style='font-family: times new roman,new york,times,serif; font-size: 12pt; color: #000000'>Dear Rita,<div>It seems like any of the suggested protocols may work, because we have done several transfections of <i>L. braziliensis </i>using the 450V 500uF protocol in the lab (Kapler <i>et al</i> 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.</div><div>Have you checked the IC50 of G418 for your specific strain? We have found it is dependent on strain and batch used.</div><div><br></div><div>All the best,</div><div><br></div><div><br><div><span name="x"></span> Tiago Rodrigues Ferreira<br> Laboratório de Parasitologia Molecular <br> Faculdade de Medicina de Ribeirão Preto <br> Universidade de São Paulo <br> Av. Bandeirantes, 3900 - CEP 14049-900<br> Ribeirão Preto - SP - Brasil <br> +55 (16) 3602 3053 <span name="x"></span><br></div><br><hr id="zwchr"><blockquote style="border-left:2px solid rgb(16, 16, 255);margin-left:5px;padding-left:5px;color:#000;font-weight:normal;font-style:normal;text-decoration:none;font-family:Helvetica,Arial,sans-serif;font-size:12pt;"><b>De: </b>"Rita Mukhopadhyay" <rmukhop@fiu.edu><br><b>Para: </b>"Patrick Bastien" <patrick.bastien@univ-montp1.fr>, leish-l@lineu.icb.usp.br<br><b>Enviadas: </b>Terça-feira, 18 de Fevereiro de 2014 3:40:44<br><b>Assunto: </b>Re: [Leish-l] S.O.S.<br><br>Dear Colleagues,<br><br>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.<br><br>Thanks a lot for your help.<br><br>Rita Mukhopadhyay<br>_______________________________________________<br>Leish-l mailing list<br>Leish-l@lineu.icb.usp.br<br>http://lineu.icb.usp.br/cgi-bin/mailman/listinfo/leish-l<br><br>--<br>This email was sent by icb.usp.br<br></blockquote><br></div></div></body></html>