RT-PCR of highly coiled RNA

[Vince Mulholland]

I am interested in this thread as it seems we have a similar problem. Ours
relates to the RT-PCR detection of plant viroids.

A viroid is small (about 360 bases), and consists of a covalently-closed
circle of single-stranded RNA. This nucleic acid has a high degree of
secondary structure. Some isolates of the viroid are refractory to RT-PCR,
and we presume that this is due to problems at the RT step. To minimise
these problems we perform the RT at 50oC.

I was wondering if there was anyone out there who was working on plant
viroids who had looked at this problem?

Vince

Vincent Mulholland,
Senior Plant Pathologist,
Diagnostics & Molecular Biology Section,
Scottish Agricultural Science Agency,
East Craigs,
Edinburgh EH12 8NJ,
U.K.

URL:    http://www.sasa.gov.uk/
E-Mail: Vince.Mulholland at sasa.gsi.gov.uk

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