Probe/primer details
| MS1414 | Tested for in situ hybridization. |
| Accession no. | pB-1146 |
| Taxonomy | Methanosarcinales; Methanomicrobia; Euryarchaeota; Archaea |
| Specificity | Methanosarcina, Methanococcoides, Methanolobus |
| Category | methanogenic archaea |
| Target rRNA | 16S rRNA |
| Position | 1414-1434 |
| Sequence | 5'- CTC ACC CAT ACC TCA CTC GGG -3' |
| G+C content [%] | 62 |
| Length [nt] | 21 |
| Check specificity/coverage | 
|
| Formamide [%] | 50 |
| Hybridization efficiency |  |
| References | Group-specific 16S rRNA hybridization probes to describe natural communities of methanogens. Raskin L, Stromley JM, Rittmann BE, Stahl DA. Applied and environmental microbiology. 1994. Pubmed An update and optimisation of oligonucleotide probes targeting methanogenic Archaea for use in fluorescence in situ hybridisation (FISH). Crocetti G, Murto M, Björnsson L. Journal of microbiological methods. 2006. Pubmed |
| Remarks | helper nucleotide hMS1395: ggtttgacgggcggtgtg helper nucleotide hMS1480: cgacttaaccccccttgc |
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).
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.
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
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.
