Extracting DNA from submerged pine wood.
A DNA extraction protocol for submerged pine logs was developed with the following properties: (i) high molecular weight DNA, (ii) PCR amplification of chloroplast and nuclear sequences, and (iii) high sequence homology to voucher pine specimens. The DNA extraction protocol was modified from a cetyltrimehtylammonium bromide (CTAB) protocol by adding stringent electrophoretic purification, proteinase K, RNAse, polyvinyl pyrrolidone (PVP), and Gene Releaser. Chloroplast rbcL (ribulose-1,5-bisphosphate carboxylase) could be amplified. Nuclear ribosomal sequences had >95% homology to Pinus taeda and Pinus palustris. Microsatellite polymorphism for PtTX2082 matched 2 of 14 known P. taeda alleles. Our results show DNA analysis for submerged conifer wood is feasible.
Reynolds
M Megan
MM
Graduate Genetics Program, Texas A&M University, College Station, TX 77844, USA.
Williams
Claire G
CG
D002467:Cell Nucleus / Q000235:genetics; D000077286:Cetrimonium; D002593:Cetrimonium Compounds / Q000737:chemistry; D002736:Chloroplasts / Q000235:genetics*; D018744:DNA, Plant / Q000235:genetics / Q000302:isolation & purification*; D019286:Endopeptidase K / Q000737:chemistry; D018895:Microsatellite Repeats / Q000235:genetics*; D028223:Pinus; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D011205:Povidone / Q000737:chemistry; D012260:Ribonucleases / Q000737:chemistry; D012270:Ribosomes / Q000235:genetics*; D012273:Ribulose-Bisphosphate Carboxylase / Q000235:genetics; D014934:Wood
D002593:Cetrimonium Compounds; D018744:DNA, Plant; D012260:Ribonucleases; D019286:Endopeptidase K; D012273:Ribulose-Bisphosphate Carboxylase; D011205:Povidone; D000077286:Cetrimonium