Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenies. PCR Protocols, A Guide to Methods and Applications . M. A. Innis, D. H. Gelfand, J. J. Sninsky and T. J. White. Obtain all stocks and place them on ice. Use gloves at all times.
Absolute cleanliness is necessary.
Set up the information in your data book. Prepare a master mix of everything except DNA and TAQ polymerase 4. Aliquot 48 ul to each PCR tube. Add 1 ul DNA to each tube using ART tips and changing tips between each sample 6. Remove samples from heat and add 1 ul Taq polymerase to each tube.