Tryptose phosphate broth solution

Manufactured by Merck Group

Sourced in United States

Tryptose phosphate broth solution is a culture medium used for the cultivation and isolation of microorganisms. It provides the necessary nutrients and growth factors for the growth and development of a wide range of bacteria, yeasts, and fungi. The solution contains tryptose, which serves as a source of nitrogen, and phosphates to maintain the appropriate pH range for microbial growth.

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Lab products found in correlation

Penicillin streptomycin, by Thermo Fisher Scientific (11 mentions) Penicillin streptomycin, by Merck Group (5 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions) Leibovitz s l 15 medium, by Thermo Fisher Scientific (4 mentions) Non essential amino acids (neaa), by Thermo Fisher Scientific (4 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (3 mentions) L glutamine, by Merck Group (3 mentions) Fetal bovine serum (fbs), by GE Healthcare (3 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (3 mentions) Indubiose, by MP Biomedicals (2 mentions) Streptomycin, by Merck Group (2 mentions) Penicillin, by Merck Group (2 mentions) L glutamine, by Thermo Fisher Scientific (2 mentions) Fetal bovine serum (fbs), by Merck Group (2 mentions) Non essential amino acids (neaa), by Merck Group (2 mentions) Non enzymatic cell dissociation solution, by Merck Group (2 mentions) Cytofix cytoperm kit, by BD (2 mentions) Penicillin, by Thermo Fisher Scientific (2 mentions) High glucose m199 medium, by Thermo Fisher Scientific (2 mentions) Dulbecco s modified eagle medium dmem, by Thermo Fisher Scientific (2 mentions)

Spelling variants of this product

Tryptose phosphate broth (121 citations) Tryptose phosphate (10 citations) Tryptose (4 citations)

22 protocols using tryptose phosphate broth solution

Isolation and Culture of Avian Muscle Cells

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QM-7 cell culture: QM-7 cells were cultured in M199 (Gibco, Gaithersburg, MD, USA) supplemented with 10% fetal bovine serum (Hyclone, Logan, UT, USA), 10% tryptose phosphate broth solution (Sigma Life Science, St. Louis, MO, USA) and 0.2% penicillin/streptomycin (Invitrogen, Carlsbad, CA, USA). The differentiation of QM-7 was induced by M199 supplemented with 10% tryptose phosphate broth solution (Sigma) and 0.2% penicillin/streptomycin.
CPM isolation and culture: The leg muscle of E11 chickens were used to isolate CPM. When the skin and bones removed, the leg muscles were rapidly chopped into small pieces in petri dish containing DMEM (Gibco) media supplemented with 20% fetal bovine serum (Hyclone) and 0.2% penicillin/streptomycin. The muscle suspending liquid was shaken by vortexing 1 min and filtered to obtain single cell. Repeat the vortexing and filtering steps for 4–6 times to release enough cells. The cells were then collected by centrifugation at 1000 × g for 5 min at room temperature. Decant medium and discard. Add DMEM supplemented with 20% FBS and 0.2% penicillin/streptomycin to resuspend cells completely. Serial plating was needed to remove fibroblasts and enrich myoblasts. The differentiation of myoblasts was induced by DMEM supplemented with 0.2% penicillin/streptomycin.

Li G., Luo W., Abdalla B.A., Ouyang H., Yu J., Hu F., Nie Q, & Zhang X. (2017). miRNA-223 upregulated by MYOD inhibits myoblast proliferation by repressing IGF2 and facilitates myoblast differentiation by inhibiting ZEB1. Cell Death & Disease, 8(10), e3094-.

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Plaque Assay for Virus Titration

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Confluent BHK-21 cell monolayers were infected with 10-fold serial dilutions of virus stock, overlaid with Eagle overlay media supplemented with 5% tryptose phosphate broth solution (Sigma Aldrich), penicillin (100 units/ml) and streptomycin (100 μg/ml) (Sigma Aldrich) and 0.6% Indubiose (MP Biomedicals) and incubated for 48 hours at 37°C. Cells were fixed and stained with 1% (w/v) methylene blue in 10% (v/v) ethanol and 4% formaldehyde in PBS.
Fixed plaques were scanned, and images measured using a GNU Image Manipulation Program IMP (GIMP, available at https://www.gimp.org). For each plaque, horizontal and vertical diameter in pixels was taken and an average of these two values was calculated. All plaques per well were measured.

Ward J.C., Lasecka-Dykes L., Neil C., Adeyemi O.O., Gold S., McLean-Pell N., Wright C., Herod M.R., Kealy D., Warner E., Jackson T., King D.P., Tuthill T.J., Rowlands D.J, & Stonehouse N.J. (2022). The RNA pseudoknots in foot-and-mouth disease virus are dispensable for genome replication, but essential for the production of infectious virus. PLoS Pathogens, 18(6), e1010589.

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Culturing Baby Hamster Kidney Cells

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Baby Hamster Kidney-21 cells (BHK-21 cells, fibroblasts) ([C-13], ATCC) were cultured in Glasgow’s Minimum Essential Medium (GMEM) (Thermofischer Scientific) supplemented with 10% (v/v) Fetal Bovine Serum (FBS) (Biowest, Nuaillé, France) and Tryptose Phosphate Broth solution (Sigma-Aldrich) at 37 °C and 5% CO₂.

Lannes N., Garcia-Nicolàs O., Démoulins T., Summerfield A, & Filgueira L. (2019). CX3CR1-CX3CL1-dependent cell-to-cell Japanese encephalitis virus transmission by human microglial cells. Scientific Reports, 9, 4833.

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Culturing Mammalian and Insect Cell Lines

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Baby Hamster Kidney cells (BHK-21/J, kindly provided by Charles M. Rice, Rockefeller University, New York, NY, USA) were grown in MEM (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) containing 7.5% FBS (Sigma, St. Louis, MO, USA), 1% L- glutamine and 1% non-essential amino acids (NEAA) (Gibco). Vero B4 cells were cultured in DMEM (Gibco) containing 1% L-glutamine and 10% FBS. Leibovitz’s L-15 Medium (Gibco) with 10% FBS (Sigma, St. Louis, MO, USA) was used to grow C6/36 cells (ATCC). Aag2 cells (CCL/FLI, Greifswald, Germany) were cultured in Leibovitz’s L-15 Medium (Gibco) supplemented with 10% FBS (Sigma, St. Louis, MO, USA), 10% tryptose phosphate broth solution (Sigma, St. Louis, MO, USA) and 1% NEAA. BHK and Vero cells were kept at 37 °C with 5% CO₂. The insect cells were cultured at 28 °C without CO₂.

Karliuk Y., vom Hemdt A., Wieseler J., Pfeffer M, & Kümmerer B.M. (2021). Characterization and Vector Competence Studies of Chikungunya Virus Lacking Repetitive Motifs in the 3′ Untranslated Region of the Genome. Viruses, 13(3), 403.

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LAMP1 Knockout Cells Infection Analysis

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LAMP1 knock-out cells (gift of S. Whelan, Harvard Medical School) were grown in IMDM-GlutaMAX media supplemented with 10% FBS and 100g/mL penicillin/streptomycin. BHK-21 cells were grown in DMEM supplemented with 10% FBS, 100g/mL penicillin/streptomycin, 2mM L-glutamine, 7% tryptose phosphate broth solution (Sigma), and 0.56% glucose (wt/vol). LAMP1 knock-out cells and BHK control cells were collected with non-enzymatic cell dissociation solution (Sigma) and incubated with LCMV at a MOI of 1 at 37°C for 1 hour. Cells were washed twice with PBS and plated in their respective media. Cells were collected 24 hours post-infection, fixed, permeabilized (BD cytofix/cytoperm kit), and stained with Alexa Fluor 488 conjugated LCMV anti-NP (clone VL-4 ^{71 (link)}) antibody. Cells were washed and analyzed by flow cytometry (LSR II, Becton Dickinson) and data subsequently analyzed with FlowJo software (FlowJo, LLC).

Hastie K.M., Igonet S., Sullivan B.M., Legrand P., Zandonatti M.A., Robinson J.E., Garry R.F., Rey F.A., Oldstone M.B, & Saphire E.O. (2016). Crystal structure of the prefusion surface glycoprotein of the prototypic arenavirus LCMV. Nature structural & molecular biology, 23(6), 513-521.

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Cell Culture Protocols for Arbovirus Research

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Aag2 cells were cultured at 25°C in Leibovitz L-15 medium (Gibco) supplemented with 10% heat inactivated fetal calf serum (FCS; PAA), 2% tryptose phosphate broth solution (Sigma), 1x MEM non-essential amino acids (Gibco), and 50 U/ml penicillin and 50 μg/ml streptomycin (pen/strep; Gibco). U4.4 and C6/36 were kept in the same culture medium at 28°C. BHK-21 cells were cultured at 37°C, 5% CO₂ in Dulbecco’s modified Eagles medium (DMEM) supplemented with 10% FCS and pen/strep. Stocks of DENV serotype 2 (DENV2), New Guinea C (NGC) and 16681 strains were grown on C6/36 cells and titred on BHK-15 cells as detailed in [40 (link)].

Miesen P., Ivens A., Buck A.H, & van Rij R.P. (2016). Small RNA Profiling in Dengue Virus 2-Infected Aedes Mosquito Cells Reveals Viral piRNAs and Novel Host miRNAs. PLoS Neglected Tropical Diseases, 10(2), e0004452.

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Isolation and Transfection of Chicken Myoblasts

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Chicken primary myoblasts were isolated from the leg muscles of E11 chickens. Leg muscle (1 g) was minced into sections of approximately 1 mm with scissors and digested with 0.25% trypsin (Gibco, Grand Island, NY, USA) at 37 °C in a shaking water bath (90 oscillations/min). Digestions were terminated by adding foetal bovine serum (Gibco) after 30 min. The mixture was filtered through a nylon mesh with 70 µm pores (BD Falcon). The filtered cells were centrifuged at 350g for 5 min, and maintained in Dulbecco’s modified Eagle medium (DMEM) (Gibco), supplemented with 20% foetal bovine serum, and 0.2% penicillin/streptomycin (Invitrogen, Carlsbad, CA, USA) at 37 °C in a 5% CO₂, humidified atmosphere. Serial plating was performed to enrich myoblasts and remove fibroblasts. DF-1 cells were cultured in DMEM with 10% foetal bovine serum and 0.2% penicillin/streptomycin. QM-7 cells were cultured in high-glucose M199 medium (Gibco) with 10% foetal bovine serum, 10% tryptose phosphate broth solution (Sigma, Louis, MO, USA) and 0.2% penicillin/streptomycin. Cells were transfected with 50 nM of miRNA mimics, 100 nM of miRNA inhibitors, 100 nM of siRNA or plasmid (1 µg/ml) using Lipofectamine 3000 reagent (Invitrogen) according to the manufacturer’s instructions.

Ouyang H., Chen X., Wang Z., Yu J., Jia X., Li Z., Luo W., Abdalla B.A., Jebessa E., Nie Q, & Zhang X. (2017). Circular RNAs are abundant and dynamically expressed during embryonic muscle development in chickens. DNA Research: An International Journal for Rapid Publication of Reports on Genes and Genomes, 25(1), 71-86.

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Culturing Aedes aegypti Aag2 Cells

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Aedes aegypti Aag2 cells were cultured in Leibovitz's L-15 medium (Invitrogen) supplemented with 10% heat-inactivated Fetal Bovine Serum (PAA Laboratories), 2% Tryptose Phosphate Broth Solution (Sigma Aldrich), 1x MEM Non-Essential Amino Acids (Invitrogen) and 50 U/ml penicillin/ streptomycin (Invitrogen) at 25 °C. Aag2 cells are a widely-used non-clonal cell line of probably embryonic origin^{26 (link)} that expresses all somatic PIWI proteins and produces both primary and secondary piRNAs via ping-pong dependent amplification^{27 (link),28 (link)}. For all experiments, cells were seeded the day before and used at 70-80% confluency. Cells were regularly confirmed to be negative for mycoplasma contamination. The cell line was a gift from R. Andino, University of California, San Francisco.

Halbach R., Miesen P., Joosten J., Taşköprü E., Rondeel I., Pennings B., Vogels C.B., Merkling S.H., Koenraadt C.J., Lambrechts L, & van Rij R.P. (2020). A satellite repeat-derived piRNA controls embryonic development of Aedes. Nature, 580(7802), 274-277.

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Propagation of Bluetongue Virus Strains

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The parental virus strains used were obtained from the dsRNA virus collection at IAH Pirbright (see: www.reoviridae.org/dsRNA_virus_proteins/ReoID/BTV-isolates.htm). BTV-26 [KUW2010/02] was originally isolated from the blood of a sheep from Kuwait as previously described [43 (link)]. BTV-1 [RSArrrr/01] is the reference strain of BTV serotype 1, originally from South Africa.
Viruses were routinely propagated in BHK-21 cells (Baby hamster kidney, clone 13), obtained from the European Collection of Animal cell Cultures (ECACC– 84100501) in Glasgow MEM medium (Invitrogen) with 10% heat-inactivated foetal calf serum (FCS). BSR cells, a clone of BHK-21 cells [48 (link)], were grown in Dulbecco’s modified Eagle medium (DMEM) containing 5% FCS. KC cells, derived from C. sonorensis midges [49 (link)], were grown in Schneider’s medium containing 10% FCS with penicillin (100 U/ml) /streptomycin (100 μg/ml) and fungizone (2 μg/ml). Aedes albopictus (mosquito) C6/36 cells [50 (link)] were grown in Leibovitz’s L15 medium (PAA) with 10% FCS, 10% tryptose phosphate broth solution (Sigma T8159), and penicillin/streptomycin. Mammalian cells were incubated at 37°C in air with 5% CO₂, and insect cells were incubated at 28°C.

Pullinger G.D., Guimerà Busquets M., Nomikou K., Boyce M., Attoui H, & Mertens P.P. (2016). Identification of the Genome Segments of Bluetongue Virus Serotype 26 (Isolate KUW2010/02) that Restrict Replication in a Culicoides sonorensis Cell Line (KC Cells). PLoS ONE, 11(2), e0149709.

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Culture and Differentiation of Avian Myoblasts

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Quail muscle clone 7 cell lines were cultured in M199 medium (Gibco, United States) with 10% fetal bovine serum (FBS) (Gibco, United States), 10% tryptose phosphate broth solution (Sigma Life Science, United States), and 0.5% penicillin/streptomycin (Invitrogen, United States).
DF-1 cell lines were cultured in DMEM (Gibco, United States) with 10% FBS and 0.5% penicillin/streptomycin.
Chicken primary myoblast were isolated from the leg muscle tissues of E11 chickens. The muscle tissues were digested with trypsin for 10 min after the skin and bones were removed. After neutralization with complete medium, the cells were then collected by centrifugation at 1000 × g for 5 min. The differential attachment was used here to eliminate fibroblasts. Growth medium (GM) for primary myoblasts contained Roswell Park Memorial Institute (RPMI)-1640 medium (Gibco, United States) with 20% FBS and 0.5% penicillin/streptomycin. To induce myogenic differentiation, GM was replaced by differentiation medium (DM) containing PRMI-1640 with 5% FBS and 0.5% penicillin/streptomycin after CPM cells reached 80∼90% confluence.

Wang Z., Ouyang H., Chen X., Yu J., Abdalla B.A., Chen B, & Nie Q. (2018). Gga-miR-205a Affecting Myoblast Proliferation and Differentiation by Targeting CDH11. Frontiers in Genetics, 9, 414.

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