PRODUCTION OF RELIABLE RANDOMLY AMPLIFIED POLYMORPHIC DNA (RAPD) MARKERS FROM DNA OF WOODY-PLANTS

A procedure for identifying reproducible RAPD markers from woody plant DNA is presented. The procedure relies on using a PCR buffer that contains 1 % Triton-X-100 and 0.1% gelatin [previously described for successful polymerase chain reaction (PCR) amplification of 16S/23S rRNA intergenic spacer regions from eubacteria], and amplification conditions of 50 cycles: 30 sec at 94C, 70 sec at 48C, and 120 sec at 72C. The combination of this buffer and these conditions amplified consistent fragments in higher amounts, as compared to other standard PCR buffers and conditions generally used for RAPD analysis. This procedure resulted in reliable RAPD patterns for all organisms tested. Chemical name used: alpha-[4-(1,1,3,3,-tetramethylbutyl)phenyl]-omega-hydroxypoly(oxy-1,2-ethanediyl) (Triton-X-100).