Description
This 47-page manual includes the agenda for the Phytophthora diagnostics workshop and the lab protocols for Phytophthora diagnostic tools.
List of lab protocols:
- Field collection of Phytophthora
- Isolation and morphological observations of unknown
species - Use Lucid Key to identify unknown species
- Use US and NL Phytophthora database and NCBI
sequence to identify unknown species - DNA extraction – Direct PCR from fresh/dried tissue
(PINF/ITS) - DNA extraction methods: simple to complex and kits
- PCR-RFLP of ITS amplicons
- Digest PCR-RFLP of ITS amplicons; Run gel for the ITS
PCR products from Lab5 - Phytophthora identification with TaqMan real time PCR
- Run gels for PCR-RFLP of ITS amplicons from Lab8
- Pack Lock Probe (PLP): Ligation & Exonuclease
- PLP: SYBRGreen PCR of ligated samples; PCR-RFLP
analysis for Lab10 - PLP: Hybridizations
- Novel detection methods
- PLP: Stain microarray and Interpretation; “Unknown
species revealed” to students