probeBase 2016
Published on probeBase 2016 (https://probebase.csb.univie.ac.at)

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Probe/primer details

Rfla729 Tested for in situ hybridization.
Full name
(Alm et al. 1996)
S-*-Rfla-0729-a-A-18
Accession no. pB-557
Taxonomy Ruminococcus [1]; Ruminococcaceae [1]; Clostridiales [1]; Clostridia [1]; Firmicutes [1]; Bacteria [1]
Specificity Ruminococcus albus, R. flavefaciens
Competitor 5'-AAA GCC CAG TAG GCC GCC -3'
5'-AAG GCC CAG TAA GCC GCC -3'
Target rRNA 16S rRNA
Position 729-746
Sequence 5'- AAA GCC CAG TAA GCC GCC -3'
G+C content [%] 61
Length [nt] 18
Check specificity/coverage [2]
Formamide [%] 20
Hybridization efficiency [2]
References

Extensive set of 16S rRNA-based probes for detection of bacteria in human feces. Harmsen HJ, Raangs GC, He T, Degener JE, Welling GW. Applied and environmental microbiology. 2002. Pubmed [2]

Design and validation of 16S rRNA probes to enumerate members of the Clostridium leptum subgroup in human faecal microbiota. Lay C, Sutren M, Rochet V, Saunier K, Doré J, Rigottier-Gois L. Environmental microbiology. 2005. Pubmed [2]

Remarks 15 min treatment with 10 µl of 1 mg lysozyme per ml in 100 mM Tris-HCl (pH 8.5). More competitors: 3. AAAGCCCAGCAAGCCGCC and 4. TCAGTCCAGTAAGCCGCC.

Print [1]

Glossary
Name (Alm et al., 1996). Probe designation according to Alm, E. W., Oerther, D. B., Larsen, N., Stahl, D. A., Raskin, L. (1996). The oligonucleotide probe database. Appl Environ Microbiol 62: 3557-9. Abstract (PUBMED) [2].
Position. Probe position according to the E. coli gene numbering.
Sequence. Sequence in IUPAC code: R=G/A, Y=T/C, M=A/C, K=G/T, S=G/C, W=A/T, H=A/C/T, B=G/T/C, V=G/C/A, D=G/A/T, N=G/A/T/C
Tm. Dissoziation temperature according to: Tm=64.9 + 41 x ((G + C - 16.4)/length).
Hybridization efficiency. Use this tool to assess in silico sensitivity (i.e. the hybridization efficiency of the oligonucleotide with its fully complementary target sequence, calculated with ProbeMelt [2].
Formamide. Percent formamide in the hybridization buffer for optimal hybridization conditions in FISH experiments.
Coverage. Coverage of the three domains calculated using the SILVA reference database 106 if no or a single mismatch is allowed. The detailed method is described in Klindworth et al., 2012. Nucleic Acids Res. 10.1093/nar/gks808 Full Text [2]
Check specificity/coverage. Use these options to reveal the in silico specificity (i.e. number of matching rRNA sequences outside the target taxon) and coverage (i.e. percentage of matching rRNA sequences within the target taxon) of an oligonucleotide against the most recent SSU and LSU rRNA sequence databases.

probeBase by Daniel Greuter, Alexander Loy, Matthias Horn, and Thomas Rattei, Department of Microbiology and Ecosystem Science, University of Vienna
Questions, comments, bug reports, and suggestions are welcome at probebase@microbial-ecology.net
Greuter et al. 2016. Nucleic Acids Res. 10.1093/nar/gkv1232

Imprint

Source URL:https://probebase.csb.univie.ac.at/pb_report/probe/557

Links
[1] https://probebase.csb.univie.ac.at/https [2] https://probebase.csb.univie.ac.at/http