Protocols in Diagnostic and Clinical Mycology
Edited by: Bert Gerrits Gerrits van den Ende (Centraalbureau voor Schimmelcultures, Utrecht, The Netherlands) Email and Carol Munro (University of Aberdeen, Aberdeen, Scotland) Email.
DNA Extraction
- Extraction of ribosomal DNA by mechanical lysis
- Extraction of ribosomal DNA by Möller (modified)
- Extraction with FastPrep kit (Bio 101)
Purification
- Quantification of nucleic acid, RNA and oligonucleotides
- Purification of PCR products by means of Microspin columns
- Purification of DNA with GeneClean kit II by means of glassmilk
- Purifying rDNA or amplicons in solution or agarose with GFX-columns
Molecular Techniques
- Executing a PCR with ribosomal DNA
- Performing ARDRA with PCR-rDNA
- Preparing an agarose-gel for PCR and ARDRA
- Sequencing using capillary electrophoresis on the ABI Prism 310
- Sequencing using 96-capillary electrophoresis on the ABI Prism 3700
- Fingerprinting: M13 RAPD
- Performing AFLP experiments on the ABI Prism 310
- Fluorescent in situ hybridization with fungal material
- Non-radioactive single-strand conformation polymorphism (SSCP) of PCR products
- Cloning of PCR amplicons in competent cells using Invitrogen TOPO® TA Cloning kit
