From field to film: rapid sequencing methods for field-collected plant species.
Methods that have proven effective in the rapid extraction of plant DNA, PCR amplification and sequencing of ribosomal and chloroplast genes are presented. Techniques that can be used under field conditions to preserve DNA for subsequent extraction are reviewed. Tissues that are fresh, heat-desiccated, silica gel-desiccated and cetyltrimethylammonium bromide (CTAB) buffer preserved are compared for DNA quality and quantity. The "delayed CTAB" method yields high molecular weight DNA, thereby providing an alternative to preservation using silica gel. Optimized methodologies for PCR amplification and double-stranded sequencing of the product are detailed including gel purification of PCR products using DEAE membranes.
Nickrent
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Southern Illinois University, Carbondale.
D000077286:Cetrimonium; D002593:Cetrimonium Compounds; D002736:Chloroplasts / Q000737:chemistry; D004247:DNA / Q000737:chemistry*; D003890:Desiccation; D008970:Molecular Weight; D010944:Plants / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012270:Ribosomes / Q000737:chemistry; D017422:Sequence Analysis, DNA / Q000379:methods*; D014021:Tissue Preservation
D002593:Cetrimonium Compounds; D004247:DNA; D000077286:Cetrimonium