Results I. Species-specific primer design

A. Design primer for Rhizoctonia solani

1) Sequence of primer PITS2RSc:

   3' AGT GGA ACC AAG CAT AAC ACT GAG 5'

2) Primer properties

a) Melting Point = 70ºC

b) Length = 24 nucleotides

c) Length of framed sequence = ~510 base pairs

B. Design primer for Sclerotinia homoeocarpa

1) Sequence of primer PITS2SHc:

   3' GGC GAC CCA GGA CTT GCA TCA TTG TG 5'

2) Primer properties

a) Melting Point = 82ºC

b) Length = 26 nucleotides

c) Length of framed sequence = 438 base pairs
II. Designed primer tests

A. Positive controls used for primer tests

B. Rhizoctonia solani

1) Initial primer test - Perkin-Elmer PCR machine

2) Primer test - Rapid Cycler PCR machine

3) Primer test on infected turfgrass

4) Primer is genus-specific, not species-specific.

C. Sclerotinia homoeocarpa

1) Initial primer test - Perkin-Elmer PCR machine

2) Primer test - Rapid Cycker PCR machine

3) Primer is not specific to S. homoeocarpa - Rapid Cycler PCR machine
III. Reduced time for diagnosis

A. A rapid DNA method was found

B. Rapid Cycler PCR machine takes 3 hours less than machine used previously.