I am currently establishing protocols for identification of plant pathogenic species by PCR. I tried primer pair BC108/BC563 published in 2006 by Rigottii and collegues, but it does not work with me (while I get excellent amplification with ITS1/ITS4 primer pair). Any suggestions or any experiences with BC108/BC563 ?
Hey Andreas,
ITS1 and ITS4 worked in many cases in our lab. Why would you use another primer pair when these two work great?
For the identification of Botrytis you could also use AEP26 and AEP27 from Brouwer et al., 2003 (in FEMS). This primer pair is supposed to target actin and yields a 233 bp fragment. Annealing is at 62°C.
It is irritating but happened to me a lot too - published primer pairs that do not work in another lab...
Good luck!
hello,
can anyone please recommend primers to detect all species of Botrytis at the same time?
Hei,
To obtain a sensitive and specific assay, I would recommend the use of a primer/probe -based real-time PCR assay. It is probably very challenging to use the ITS rDNA gene cluster to design a specific assay for Botrytis cinerea. One option is to use betatubulin -see e.g. New Phytol (2005) 168: 465-474.
- Badges
- Science method