I am identifying Paecilomyces variotii isolates through a poliphasic approach. At the molecular side, I have extracted the fungal DNA from culture in YES using a protocol with PrepMan (Life).
The DNA obtained presents good quality (A260/280: >=1.8 ,A260/230: ~1.8) and is diluted to 40 ng/mL. I have already successfuly amplified and sequenced part of the ITS region from this DNA. However I haven't been able to amplify the elongation factor.
The primers I am using are EF-1H and EF-2T. I have already succesfuly sequenced the elongation factor region from others species such as Purpureocillium lilacinum using the same primer pair.
It would be of great help if anyone could give some input.
Hi Xafranski
I think it would better to use cDNA, inspite of DNA. If still problem please try with other primers http://sites.biology.duke.edu/fungi/mycolab/primers.htm
I hope it can help you.
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