Phytoplasma Plasmid DNA Extraction
Phytoplasma plasmids have generally been detected from DNA extracted from plants and insects using methods designed for the purification of total phytoplasma DNA. Methods include extraction from tissues that are high in phytoplasma titre, such as the phloem of plants, with the use of CsCl–bisbenzimide gradients that exploit the low G+C content of phytoplasma DNA. Many of the methods employed for phytoplasma purification have been described elsewhere in this book. Here we describe in detail two methods that are specifically aimed at isolating plasmid DNA.
Key wordsAlkaline lysis CsCl gradients Plasmid
- 4.Namba S, Oshima K, Gibb K (2005) Phytoplasma genomics. In: Blanchard A, Browning GS (eds) Mycoplasmas: molecular biology, pathogenicity and strategies for control. Horizon Bioscience, Norfolk, pp 97–133Google Scholar
- 11.Doyle JJ, Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus 12:13–15Google Scholar
- 20.Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual, 3rd edn. Cold Spring Harbour Press, New YorkGoogle Scholar
- 21.Ausubel FM et al (eds) (1992) Current protocols in molecular biology. Wiley, New YorkGoogle Scholar