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. 2013 Jul 30;5(7):63. doi: 10.1186/gm467

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
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