can anybody suggest the best way of increasing the specificity of Ryan 2013 nasted primers for 18S rRNA of Cryptosporidium spp detection? in other word, that primer cross reacting with other coccidia(Eimeria-Isospora) as well as eukaryotic fresh water gene?
Oľga Danišová
You can do the third reaction with more specific primer after nested PCR and thus better localize the polymorph area they need for the identification of the pathogen.
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