Primer for Unkown Protein PCR - How to design primer for unknown protein (May/18/2010 )
Good day!
I'm a noob in molecular biology. Please, jelp me guys
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
Xylph on May 19 2010, 12:05 AM said:
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
confusing here..
1. what species are u using now?
2. what u means with 'identify (only) Heat-resistant Mold (HRM) in sample'? does it means, u want to know what kind of species (HRM) contain in ur samples?
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