Protocol

Centre : Central Potato Research Institute(CPRI)
Name of the Protocol
Dection of Phytophthora infestans using SCAR marker.
Nature : Novel
Crop / Pathogen : Potato /Phytophthora infestans.

Inventors

1. Touseef Hussain
2. Mehi Lal
3. BP Singh
4. Sanjeev Sharma

Objectives

1. Detction of P. infestans from host tissues.

Procedure / Summary

The primer PITH2-F/PITH2-R was developed and used for this study. PCR reactions was performed in a total volume of 25µl, containing1X Taq DNA Buffer with 1.5mM MgCl2, 5U/µl Dream Taq DNA Polymerase, 200Mm of each dNTPs, 10 pmol of each Forward and Reverse primer, 10ng/ul template DNA, milliQ water (variable) per reaction. The PCR conditions are as follows: denaturation at 94 °C for 2 min, followed by 30 cycles of 94 °C for 30 s, 58-60 °C for 30 s, 72 °C for 1 min and a final extension at 72 °C for 10 min. Amplified PCR products (524 bp) were separated by electrophoresis on 1.8 % agarose gel and picture was captured under gel documentation system.

Applications

1. Detection of latent infection in seed lots.

Publications

1. Not published.