Primer (For PCR)

[erynna]

Hi All

 

I'm currently working in the UK designing practical classes for college students and I'm in need of some plant biotechnology advice. Would anyone be able to recommend me a universal plant primer to use in PCR? I'm not looking to amplify a specific gene or sequence but I'm having difficulties finding a primer which is appropriate. I have considered using a universal rice primer or a universal chloroplast primer.

 

I just wish to demonstrate the procedure of PCR and DNA fingerprinting to the students.

 

Erynna

[CharonY]

Well, it really depends on what you want to look at and what really you want to demonstrate. If you just want to run a PCR you could even use random primers (essentially like a RAPD, see Welsh, J. and McClelland, M., Nucleic Acids Res., 1991, 19, 6823–6831). Its use for plants is documented in: Tingey, S. V et al., RAPD Technology to Plant Breeding (ed. Neff, M.), ASHS Publishers, Minnesota, 1993, pp. 3–8 and

Williams et al., Methods Enzymol., 1993, 218, 704–740.

 

If you got the sequence of the plant(s) you want to identify it is obiously even easier (align sequence, build primers based on conserved seqs).

 

A database of potential plan mol. markers is documented by Rudd et al., Nucleic Acids Res. 2005 33(Database Issue):D628-D632

[erynna]

I thought about using random primers but I believe they would not result in clear bands. It would be nice if the students are able to see slightly different bands for the different plant species.

 

This is the paper on the use of universal rice primers. http://www.seoulin.co.kr/upload/biofiles/2003812115152.pdf

 

I have considered trying these primers out but if someone knows of a primer that will yield similar results I’m open for suggestions.

 

Thanks

Erynna