FIELD: biotechnology.
SUBSTANCE: invention represents a method for identifying Heterobasidion annosum fungus, based on PCR, involving DNA recovery from a plant substrate or fungal mycelium, real-time PCR amplification involves DNA denaturation, primer attachment and DNA chain elongation. PCR amplification involves using primers: forward SEQ ID NO: 1 GAATATCGTGCAAGGTTG and reverse SEQ ID NO: 2 CGAAGAGTTTGTGAGAAG, and also TaqMan probe SEQ ID NO: 3 FAM-CGCGGCTCGGAAGGGGTCAAAAGAACCC-BHQ1. Following temperature cycles are used: 1 cycle – temperature 94 °C for 5 minutes and 44 cycles – alternately temperature 94 °C for 20 s and temperature of 59 °C for 60 s. Logarithmic increase of the fluorescence level up to 38 cycle in the FAM channel of the thermocycler testifies to the presence of fungus Heterobasidion annosum in the analyzed sample.
EFFECT: invention enables fast identification of fungus Heterobasidion annosum.
1 cl, 1 dwg, 1 tbl, 1 ex
