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Spectinomycin sulfate

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Spectinomycin sulfate is a broad-spectrum antibiotic used in the laboratory setting. It functions as an inhibitor of bacterial protein synthesis, disrupting the translocation process during bacterial cell division. This product is commonly used in microbiological research and applications.

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Lab products found in correlation

Fusidic acid sodium salt, by Merck Group (3 mentions) Kanamycin, by GoldBio (2 mentions) Chloramphenicol, by Thermo Fisher Scientific (2 mentions) Arabinose, by Merck Group (2 mentions) Anhydrotetracycline atc, by Merck Group (2 mentions) Ampicillin sodium salt, by Thermo Fisher Scientific (2 mentions) Viomycin sulfate, by Merck Group (2 mentions) Guanosine triphosphate, by Merck Group (2 mentions) Hygromycin b, by Merck Group (2 mentions) Gdpnp, by Merck Group (2 mentions) Mono q 5 50 gl ion, by GE Healthcare (2 mentions) Gtpγs, by Merck Group (1 mentions) Pyruvate kinase, by Merck Group (1 mentions) Monoq 5 50 gl anion exchange column, by GE Healthcare (1 mentions) Myokinase, by Merck Group (1 mentions) Phosphoenolpyruvate pep, by Merck Group (1 mentions) Sodium dodecyl sulfate (sds), by Thermo Fisher Scientific (1 mentions) H3bo3, by Thermo Fisher Scientific (1 mentions) Dna ligase, by New England Biolabs (1 mentions) Chloroform, by Thermo Fisher Scientific (1 mentions)

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Spectinomycin (9 citations)

8 protocols using spectinomycin sulfate

1

Genetic Manipulation of E. coli DH10b

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E.coli DH10b (F–mcrA Δ(mrr-hsdRMS-mcrBC) ⌽80lacZΔM15 ΔlacX74 recA1 endA1 araD139 Δ(araleu) 7697 galU galK rpsL nupG λ–)41 (link) was used for genetic manipulation and characterization. Note that E. coli DH10b has the fimE/fimB and fim structural genes deleted41 (link). Cells were grown in LB miller broth (Difco, 90003-350) for functional assays and SOB (Teknova, S0210) (2% Bacto-tryptone, 0.5% Bacto yeast extract, 10 mMNaCl, 2.5 mMKCl) for cloning. Chloramphenicol (Alfa Aesar, AAB20841-14) (34 μg/ml), kanamycin (GoldBio, K-120-10) (50 μg/ml) or spectinomycin sulfate (50 μg/mL) (MP Biomedicals LLC, 158993) were supplemented where appropriate. Arabinose (Sigma Aldrich, MO, A3256), IPTG (Isopropyl β-D-1-thiogalactopyranoside) (GoldBio, I2481C25) or anhydrotetracycline (aTc) (Sigma Aldrich, 37919) was used as inducers. For Arabinose induction system 0.5% (w/v) glucose was used to reduce the leakiness of the uninduced state. Three fluorescence proteins GFPmut342 (link), mRFP143 (link) and YFP42 (link) were used as reporters in this study.
Yang L., Nielsen A.A., Fernandez-Rodriguez J., McClune C.J., Laub M.T., Lu T.K, & Voigt C.A. (2014). Permanent genetic memory with >1 byte capacity. Nature methods, 11(12), 1261-1266.
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2

Genetic Manipulation of E. coli DH10b

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E.coli DH10b (F–mcrA Δ(mrr-hsdRMS-mcrBC) ⌽80lacZΔM15 ΔlacX74 recA1 endA1 araD139 Δ(araleu) 7697 galU galK rpsL nupG λ–)41 (link) was used for genetic manipulation and characterization. Note that E. coli DH10b has the fimE/fimB and fim structural genes deleted41 (link). Cells were grown in LB miller broth (Difco, 90003-350) for functional assays and SOB (Teknova, S0210) (2% Bacto-tryptone, 0.5% Bacto yeast extract, 10 mMNaCl, 2.5 mMKCl) for cloning. Chloramphenicol (Alfa Aesar, AAB20841-14) (34 μg/ml), kanamycin (GoldBio, K-120-10) (50 μg/ml) or spectinomycin sulfate (50 μg/mL) (MP Biomedicals LLC, 158993) were supplemented where appropriate. Arabinose (Sigma Aldrich, MO, A3256), IPTG (Isopropyl β-D-1-thiogalactopyranoside) (GoldBio, I2481C25) or anhydrotetracycline (aTc) (Sigma Aldrich, 37919) was used as inducers. For Arabinose induction system 0.5% (w/v) glucose was used to reduce the leakiness of the uninduced state. Three fluorescence proteins GFPmut342 (link), mRFP143 (link) and YFP42 (link) were used as reporters in this study.
Yang L., Nielsen A.A., Fernandez-Rodriguez J., McClune C.J., Laub M.T., Lu T.K, & Voigt C.A. (2014). Permanent genetic memory with >1 byte capacity. Nature methods, 11(12), 1261-1266.
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3

Antibiotic Efficacy Analysis

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Antibiotics used in this study included Spectinomycin Sulfate (MP Biomedicals) and Ampicillin Sodium Salt (Fisher).
Sharma A, & Wood K.B. (2021). Spatial segregation and cooperation in radially expanding microbial colonies under antibiotic stress. The ISME Journal, 15(10), 3019-3033.
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4

Buffers for Single-Molecule Experiments

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All experiments were carried out in either polymix buffer A (50 mM Tris-OAc (pH 7.5), 100 mM KCl, 5 mM NH4OAc, 0.5 mM Ca(OAc)2, 5 mM Mg(OAc)2, 6 mM 2-mercaptoethanol, 0.1 mM EDTA, 5 mM putrescine and 1 mM spermidine)45 (link) or polymix buffer B (30 mM HEPES pH 7.5, 5 mM MgCl2, 50 mM NH4Cl, 5 mM 2-mercaptoethanol, 2 mM spermidine and 5 mM putrescine)46 (link). A cocktail of triplet-state quenchers (1 mM Trolox, 1 mM nitrobenzyl alcohol and 1 mM cyclooctatetraene) and an enzymatic oxygen scavenging system (protocatechuic acid (PCA)/protocatechuate-3,4-dioxygenase (PCD)) were used for smFRET experiments. Spectinomycin sulfate was purchased from MP Biomedicals. Fusidic acid sodium salt, GTP and GTPγS were from Sigma-Aldrich. GTP was further purified using a Mono Q 5/50 GL anion exchange column (GE Healthcare Life Sciences). Pyruvate kinase, myokinase and phosphoenolpyruvate (PEP) were purchased from Sigma-Aldrich. All other standard reagents were purchased from Sigma-Aldrich or VWR.
Rundlet E.J., Holm M., Schacherl M., Natchiar S.K., Altman R.B., Spahn C.M., Myasnikov A.G, & Blanchard S.C. (2021). Structural basis of early translocation events on the ribosome. Nature, 595(7869), 741-745.
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5

Antibiotic Efficacy Analysis

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Antibiotics used in this study included Spectinomycin Sulfate (MP Biomedicals) and Ampicillin Sodium Salt (Fisher).
Hallinen K.M., Karslake J, & Wood K.B. (2020). Delayed antibiotic exposure induces population collapse in enterococcal communities with drug-resistant subpopulations. eLife, 9, e52813.
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6

Detailed Microbial Culture Protocol

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Chemicals were purchased from Sigma-Aldrich (Na2EDTA·2H2O, CaCl2·2H2O, KH2PO4, vitamin B12, ZnCl2, and spectinomycin sulfate), MP Biomedicals (FeCl3·6H2O, CuSO4·5H2O, and CoCl2·6H2O), Acros (Na2MoO4·2H2O), Amresco (NaOAc, 3M, pH 5.2), Fisher Chemical (MnCl2·4H2O and Na2S2O3), and Fisher BioReagents [NaCl, MgSO4·7H2O, KCl, NaNO3, Tris base, H3BO3, kanamycin monosulfate, SDS, chloroform, saturated phenol (pH 4.3), and absolute ethanol]. Enzymes, including Q5 DNA polymerase, Taq polymerase, DNA ligase, and restriction enzymes, were purchased from New England Biolabs. DNA isolations and purifications were performed using the Zyppy Plasmid Miniprep kit, DNA Clean & Concentrator, and Zymoclean Gel DNA Recovery kit from Zymo Research. Genomic DNA was isolated using the GenElute Bacterial Genomic DNA kit (Sigma-Aldrich). All other vendors are indicated in the subsequent methods sections.
Ruffing A.M., Jensen T.J, & Strickland L.M. (2016). Genetic tools for advancement of Synechococcus sp. PCC 7002 as a cyanobacterial chassis. Microbial Cell Factories, 15, 190.
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7

Purification and Characterization of GTP

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Fusidic acid sodium salt (Sigma-Aldrich), spectinomycin sulfate (MP Biomedicals), viomycin sulfate (USP) and hygromycin B (Sigma-Aldrich) were used at the purity stated by the commercial suppliers (≥97 %). Guanosine triphosphate (GTP), guanosine diphosphate (GDP) and GDPNP were purchased from Sigma, and GTP was further purified on a Mono Q 5/50 GL ion exchange column (GE Healthcare Life Sciences).
Wasserman M.R., Alejo J.L., Altman R.B, & Blanchard S.C. (2016). Multi-perspective smFRET reveals rate-determining late intermediates of ribosomal translocation. Nature structural & molecular biology, 23(4), 333-341.
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8

Purification and Characterization of GTP

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Fusidic acid sodium salt (Sigma-Aldrich), spectinomycin sulfate (MP Biomedicals), viomycin sulfate (USP) and hygromycin B (Sigma-Aldrich) were used at the purity stated by the commercial suppliers (≥97 %). Guanosine triphosphate (GTP), guanosine diphosphate (GDP) and GDPNP were purchased from Sigma, and GTP was further purified on a Mono Q 5/50 GL ion exchange column (GE Healthcare Life Sciences).
Wasserman M.R., Alejo J.L., Altman R.B, & Blanchard S.C. (2016). Multi-perspective smFRET reveals rate-determining late intermediates of ribosomal translocation. Nature structural & molecular biology, 23(4), 333-341.
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