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Xylph's Content

There have been 2 items by Xylph (Search limited from 30-May 19)


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#71989 Primer for Unkown Protein PCR

Posted by Xylph on 21 May 2010 - 02:01 AM in PCR, RT-PCR and Real-Time PCR

i want to make primer to detect contamination of these molds (Byssochlamys fulva, Byssochlamys nivea, Neosartorya fischeri, Talaromyces macrosporus, Talaromyces flavus) in the sample. so i need specific gene / domain which is specific on those species. They arent in the same genus, but they have something in common. All of them are heat resistant (~95C), so i think they will have specific domains on their coding sequence for the heat adaptation.

same principle: use ctx gene primer to detect Vibrio cholerae in sample.

Thx for the reply
=D



#71447 Primer for Unkown Protein PCR

Posted by Xylph on 18 May 2010 - 08:05 AM in PCR, RT-PCR and Real-Time PCR

Good day! :P
I'm a noob in molecular biology. Please, jelp me guys :D

I want to design primer to identify (only) Heat-resistant Mold (HRM) in sample. But i cant find any specific proteins in HRM, because no such data in NCBI.

1. Can I design primer without knowing specific target protein?
2. Can I design primer by comparing the genome of the HRMs and find a region that similiar in all species of HRM? Is it reliable? And what tool to use?
3. Many journals use ITS regions for detection, but i think it will give positive result for all fungi. Am I right?

common HRMs: Byssochlamys fulva, Byssochlamys nivea, Neosartorya fischeri, Talaromyces macrosporus, Talaromyces flavus
maybe someone have made primers for these HRMs before

Many thanks in advance :lol:




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