Table 1.
Summary of culture-independent methods for studying the mycobiome
Method | Procedure | Strength | Weakness |
---|---|---|---|
RFLP [10,60,61] | 1. PCR of rDNA 2. Build the clone library 3. Digest with endonucleases 4. Run capillary electrophoresis |
Allow comparisons of fungal abundance across groups | 1. Considerable intraspecific variability 2. Not specific enough to differentiate fungi at the level of species 3. Unable to quantify the proportion of each type of fungi in the mycobiome |
OFRG [62] | 1. PCR of rDNA 2. Build the clone library 3. Hybridize with oligonucleotide probes |
||
DGGE [14] | 1. PCR of rDNA 2. Build the clone library 3. Run the denaturing gel electrophoresis 4. Analyze the patterns of the bands |
||
In situ hybridization [14] | 1. Process biopsy sample 2. Probe hybridization |
||
Sanger sequencing [50] | 1. PCR of rDNA 2. Build the clone library 3. Sanger sequencing |
Specific enough to differentiate between species | High cost [63] |
Pyrosequencing [18,64] | 1. PCR of rDNA 2. Pyrosequencing |
1. Homo-polymerization 2. Environmental contamination |