A Renewed DNA Extraction Method for Molecular Study of Melampsora larici-populina

Based on the bi-ball method of DNA extraction, the study renews some steps and component, which makes the PCR more sensible. The changes include: (1) Put the urediniospores into the tube with two steel balls and 1.6 g/L Chelex-100, the uredinia may be picked up with airing host tissues. (2) Add KOH into the extraction buffer instead of NaOH and vortex the tube violently for 30 min. (3) Prick a hole under the bottom instead of picking out of the steel balls, then centrifuge the tube to toss the liquid into the sheathed tube. The DNA achieved by this way can be used as substrates of ITS-nrDNA-PCR, RAPD, SSR. Futher dilution multiples of the extraction liquid is also tested. RAPTD and SSR may complete well with over 16×DNA extraction buffer.