I have a research to find the resistance gene of Trypanosoma evansi to diminazyne drugs.
Well, there are two type of mthode that my supervisor offered to me, by using the RFLP-PCR or RT-PCR, but hosnestly I never know about these two methods, so could anyone here can help me which one the best methods to find the resistance gene of Trypanosoma?
Could you share the journal or link to read.
I am looking forward your sharing
Regards,
hi see this link
https://www.ncbi.nlm.nih.gov › NCBI › Literature ›
Hi M. Mirza Nuryady
You can use a genetic screening using increasing concentrations of the drug to generate drug-resistant parasites, which genome afterwards you must analyse by NGS to identify the mutation conferring resistance. This approach would be useful, but you would need to be aware that potentially you would also find a mutation of drug-related transporters. So, a double confirmation would be necessary using specific RNAi cell lines.
Furthermore, you can do a genome-scale RNAi screens for high-throughput phenotyping in bloodstream-form African trypanosomes, as described in http://www.nature.com/nprot/journal/v10/n1/full/nprot.2015.005.html
Dear Alejandro,
Thank you so much for your kind advise, could you give me some enzyme to get the resistance gene without genome scale RNAi screening before?
Thank you so much
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