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MBD0035

Sigma-Aldrich

Bacterial negative control FISH probe - Cy3

Probe for fluorescence in situ hybridization (FISH),20 μM in water

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Quality Level

200

technique(s)

FISH: suitable

fluorescence

λex 550 nm; λem 570 nm (Cy3)

shipped in

dry ice

storage temp.

−20°C

General description

Fluorescent In Situ Hybridization technique (FISH) is based on the hybridization of fluorescent labeled oligonucleotide probe to a specific complementary DNA or RNA sequence in whole and intact cells.1 Microbial FISH allows the visualization, identification and isolation of bacteria due to recognition of ribosomal RNA also in unculturable samples.2

FISH technique can serve as a powerful tool in the microbiome research field by allowing the observation of native microbial populations in diverse microbiome environments, such as samples from human origin (blood3 and tissue4), microbial ecology (solid biofilms5 and aquatic systems6) and plants7.

Prokaryotic single cell life forms are divided into two domains, called Bacteria and Archaea, originally categorized as Eubacteria and Archaebacteria.8 However both terms, Eubacteria and Bacteria are still being used in microbiology.

The Negative control non-specific, nonsense probe, is used in the FISH experiments to detect nonspecific binding of the probes and set the appropriate conditions for the experiment.9

The negative control probe was used in FISH technique on various samples such as, pure culture (as described in the figure legends) and blood cultures9,10.

It is strongly recommended to include positive and negative controls in FISH assays to ensure specific binding of the probe of interest and appropriate protocol conditions. We offer positive (MBD0032/33) and negative (MBD0034/35) control probes, that accompany the specific probe of interest.

Application

Probe for fluorescence in situ hybridization (FISH), negative control probe used to detect nonspecific binding.

Features and Benefits

  • Detection of nonspecific binding of tested probes.
  • Verification of FISH results accuracy.
  • Calibration of FISH experiment appropriate conditions.
  • Can be used to control for nonspecific binding in variuos samples such as, blood cultures and pure cultures.
  • The ability to detect bacteria in its natural habitat is an essential tool for studying host-microbiome interaction.

Storage Class Code

12 - Non Combustible Liquids

WGK

nwg

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Regulatory Information

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Dorothee Maria Gescher et al.
International journal of antimicrobial agents, 32 Suppl 1, S51-S59 (2008-08-23)
Sepsis is a life-threatening disease with a high mortality rate. Rapid identification of blood culture isolates plays a crucial role in adequate antimicrobial therapy in sepsis patients. To accelerate microbiological diagnosis, a comprehensive panel of oligonucleotide probes for fluorescence in
V A Kempf et al.
Journal of clinical microbiology, 38(2), 830-838 (2000-02-03)
Using fluorescent in situ hybridization (FISH) with rRNA-targeted fluorescently labelled oligonucleotide probes, pathogens were rapidly detected and identified in positive blood culture bottles without cultivation and biotyping. In this study, 115 blood cultures with a positive growth index as determined
C R Woese et al.
Proceedings of the National Academy of Sciences of the United States of America, 87(12), 4576-4579 (1990-06-01)
Molecular structures and sequences are generally more revealing of evolutionary relationships than are classical phenotypes (particularly so among microorganisms). Consequently, the basis for the definition of taxa has progressively shifted from the organismal to the cellular to the molecular level.
Leore T Geller et al.
Science (New York, N.Y.), 357(6356), 1156-1160 (2017-09-16)
Growing evidence suggests that microbes can influence the efficacy of cancer therapies. By studying colon cancer models, we found that bacteria can metabolize the chemotherapeutic drug gemcitabine (2',2'-difluorodeoxycytidine) into its inactive form, 2',2'-difluorodeoxyuridine. Metabolism was dependent on the expression of
Michael Wagner et al.
Current opinion in microbiology, 6(3), 302-309 (2003-07-02)
Fluorescence in situ hybridisation with rRNA-targeted nucleic acid probes can be used to directly identify microorganisms within complex samples in a few hours and therefore has widespread application in environmental and medical microbiology. The past year has seen significant methodological
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