Editing Hybridization probe (section)

=== Limitations ===
In some instances, differentiation between species may be problematic when using [[16S ribosomal RNA|16S rRNA]] sequences due to similarity. In such instances, [[23S rRNA]] may be a better alternative.<ref>{{cite journal | author1 = Fox, G.E. |author2= Wisotzkey, J.D. | author3=Jurtshuk Jr., P. | year = 1992 | title = How close is close: 16S rRNA sequence identity may not be sufficient to guarantee species identity. | journal = Int. J. Syst. Bacteriol. | volume = 42 |issue= 1 | pages = 166–170 | doi=10.1099/00207713-42-1-166|pmid= 1371061 | doi-access = free }}</ref> The global standard library of rRNA sequences is constantly becoming larger and continuously being updated, and thus the possibility of a random hybridization event between a specifically-designed probe (based on complete and current data from a range of test organisms) and an undesired/unknown target organism cannot be easily dismissed.<ref>{{cite journal | author = Olsen, G.J. |author2=Lane, D.J. |author3=Giovannoni, S.J. |author4= Pace, N.R.  |author5= Stahl, D.A. | year = 1986 | title = Microbial ecology and evolution: a ribosomal RNA approach | journal = Annu. Rev. Microbiol. | volume = 40 | pages = 337–365 | doi=10.1146/annurev.mi.40.100186.002005|pmid=2430518 }}</ref> On the contrary, it is plausible that there exist microorganisms, yet to be identified, which are phylogenetically members of a probe target group, but have partial or near-perfect target sites, usually applies when designing group-specific probes.

Probably the greatest practical limitation to this technique is the lack of available automation.<ref>{{cite journal |vauthors=Amann R, Ludwig W | year = 2000 | title = Ribosomal RNA-targeted nucleic acid probes for studies in microbial ecology | journal = FEMS Microbiology Reviews | volume = 24 | issue = 5 | pages = 555–565 | doi=10.1111/j.1574-6976.2000.tb00557.x| pmid = 11077149 | doi-access = free }}</ref>