Because of the potential for using myxobacteria as biological control agents, seven phytopathogenic prey organisms were chosen, representing a taxonomically diverse set of organisms that infect a variety of economically important plant hosts (Table 2 ). Gram-negative organisms were grown in LB broth (10 g/L tryptone (Fisher), 10 g/L NaCl, 5 g/L yeast extract) for 18–24 h at 37 °C, 180 rpm. Gram-positive and fungal organisms were grown in tryptone Soy Yeast Extract (17 g/L tryptone, 3 g/L soya peptone (Oxoid), 6 g/L yeast extract, 5 g/L NaCl, 2.5 g/L K2HPO4, 2.5 g/L glucose) or YEPS (10 g/L yeast extract, 20 g/L tryptone, 10 g/L saccharose (Fisher)), respectively, for 40 h at 30 °C, 180 rpm. Strains were maintained at 4 °C for up to two weeks on agar plates before re-plating and stored long-term as liquid glycerol stocks at −80 °C.
Tryptone
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom, United States, China, Hungary, Denmark, Germany
Tryptone is a complex organic compound derived from the enzymatic digestion of casein, a milk protein. It is commonly used as a nutrient source in microbial growth media for the cultivation of various types of bacteria. Tryptone provides a rich source of amino acids, peptides, and other essential nutrients required for the growth and proliferation of bacterial cultures in laboratory settings.
Automatically generated - may contain errors
Lab products found in correlation
Yeast extract, by Thermo Fisher Scientific (130 mentions)
Sodium chloride (nacl), by Thermo Fisher Scientific (21 mentions)
Sodium chloride (nacl), by Merck Group (21 mentions)
Glycerol, by Thermo Fisher Scientific (10 mentions)
Ampicillin, by Merck Group (9 mentions)
Kanamycin, by Merck Group (8 mentions)
Tetracycline, by Merck Group (7 mentions)
Sodium chloride (nacl), by Sinopharm (7 mentions)
Sodium chloride (nacl), by Avantor (6 mentions)
Peptone, by Thermo Fisher Scientific (6 mentions)
Isopropyl β d 1 thiogalactopyranoside (iptg), by Thermo Fisher Scientific (5 mentions)
Imidazole, by Thermo Fisher Scientific (5 mentions)
Acetonitrile, by Merck Group (5 mentions)
Chloramphenicol, by Merck Group (5 mentions)
D glucose, by Merck Group (5 mentions)
Yeast extract, by BD (4 mentions)
Sodium hydroxide (naoh), by Thermo Fisher Scientific (4 mentions)
Ampicillin, by Thermo Fisher Scientific (4 mentions)
T4 dna ligase, by New England Biolabs (4 mentions)
Imidazole, by Merck Group (3 mentions)
204 protocols using tryptone
1
Myxobacteria as Biopesticide Agents
2
Construction of S. aureus Gene Knockout Strains
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
S. aureus USA500 (Diep et al., 2006 (link)) was used for construction of gene knockout and complementation strains. E. coli DC10B (Monk et al., 2012 (link)) was used for shuttle plasmid construction. Luria Broth medium was composed of 1% tryptone (Oxoid), 0.5% yeast extract (Oxoid) and 0.5% NaCl; BM (B-Medium) was composed of 1% tryptone, 0.5% yeast extract, 0.5% glucose, 0.1% K2HPO4 and 0.5% NaCl; BM and TSB (Tryptic soy broth, Oxoid) were used for S. aureus cultivation. Bacterial strains were inoculated in BM, and their growth rate at 37°C was monitored by measuring the OD values at 600 nm. Anhydrotetracycline (ATc) was used for induction of secY antisense RNA during gene knockout. Antibiotics were added to medium at the following concentrations: chloramphenicol, 10 μg/ml; ampicillin, 100 μg/ml, levofloxacin, 50 μg/ml.
3
Interstitial Biofilm Cultivation Protocols
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Bacterial strains used in this study were the P. aeruginosa strain PAK and the Proteus vulgaris strain ATCC 13315. P. aeruginosa PAK was maintained on 1.5% LBA [10 g/L tryptone (Oxoid), 5 g/L yeast extract (Oxoid), 5 g/L NaCl, 1.5% agar (Oxoid)] and Pr. vulgaris ATCC 13315 was maintained on low salt 1.5% LBA (2.5 g/L NaCl) at 37°C. P. aeruginosa interstitial biofilms were cultured on nutrient media [4 g/L tryptone (Oxoid), 2 g/L yeast extract (Oxoid), 2 g/L NaCl, 1 g/L MgSO4.7H2O] solidified with 8 g/L gellan gum (MP Biomedicals; twitching motility gellan gum; TMGG). Pr. vulgaris interstitial biofilms were grown on LB solidified with 8 g/L gellan gum (Luria Bertani gellan gum; LBGG).
4
Quantifying Microbial Colony Formation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The colony forming units (CFU) of each experiment were determined using the spread plate method on agar. For the reactors not containing Pb(II), the agar consisted of 10 g/L tryptone (Oxoid, South Africa), 5 g/L yeast extract (Oxoid, South Africa), 10 g/L NaCl (Glassworld, South Africa), and 15 g/L agar in distilled water. For the reactors containing Pb(II), the agar consisted of 20 g/L tryptone (Oxoid, South Africa), 10 g/L yeast extract (Oxoid, South Africa), and 1 g/L NaCl (Glassworld, South Africa) in distilled water. The sample was diluted to a range of 10−4 to 10−7 in sterile distilled water. The number of colonies was counted 96 h after plating was completed.
5
Xylanase Production and Purification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Wheat arabinoxylan (WAX) was of medium viscosity and supplied by Megazyme (Wicklow, Ireland). The chemical composition of this type of WAX has previously been described [17 (link)]. Beechwood xylan (BeWX) and all chemicals were purchased from Sigma-Aldrich (St Louis, MO, USA) unless otherwise specified. AaXynA was kindly provided by the laboratory of Food Chemistry (Wageningen University) as described elsewhere [8 (link)].
Luria-Bertani (LB) medium contained per L: 10 g NaCl, 10 g tryptone (Oxoid), 5 g yeast extract (Roth). LB2 contains per liter: 10 g tryptone (Oxoid), 5 g yeast extract (Roth), 10 g sodium chloride and salts mix consisting of 1 g NH4Cl; 3 g NaCl; 1.50 g Na2SO4; 0.08 g NaHCO3; 1 g KCl; 1.8 g MgCl2 × 6H2O; 0.30 g CaCl2 × 2H2O. pH was set to 6.6 at room temperature and the medium was autoclaved for 20 min at 121 °C, after which 1 mL K2HPO4 (250 g/L) was added.
For plasmid propagation and protein expression E. coli strains DH5α and BL21(DE3) were used respectively. E. coli strains were grown in LB medium at 37 °C, pH 7.0 with an agitation speed of 150 RPM.
Geobacillus thermodenitrificans T12 was isolated from compost [18 (link)]. Strain T12 was grown in LB2 medium at 65 °C, 150 RPM and pH 7.0.
Luria-Bertani (LB) medium contained per L: 10 g NaCl, 10 g tryptone (Oxoid), 5 g yeast extract (Roth). LB2 contains per liter: 10 g tryptone (Oxoid), 5 g yeast extract (Roth), 10 g sodium chloride and salts mix consisting of 1 g NH4Cl; 3 g NaCl; 1.50 g Na2SO4; 0.08 g NaHCO3; 1 g KCl; 1.8 g MgCl2 × 6H2O; 0.30 g CaCl2 × 2H2O. pH was set to 6.6 at room temperature and the medium was autoclaved for 20 min at 121 °C, after which 1 mL K2HPO4 (250 g/L) was added.
For plasmid propagation and protein expression E. coli strains DH5α and BL21(DE3) were used respectively. E. coli strains were grown in LB medium at 37 °C, pH 7.0 with an agitation speed of 150 RPM.
Geobacillus thermodenitrificans T12 was isolated from compost [18 (link)]. Strain T12 was grown in LB2 medium at 65 °C, 150 RPM and pH 7.0.
6
Salmonella Typhimurium Preparation Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The protocol used for strain preparation (Butot et al., 2018) (link) can be summarized as follows: Salmonella enterica serovar Typhimurium SL1344 obtained as cryobeads from the LFMFP (Ghent University) laboratory collection were revived and subcultured in brain-heart infusion broth (BHI, Oxoid, England). Cells were harvested from tryptone soy agar (TSA, Oxoid, England) by adding 2 mL of sterile tryptone (Oxoid, England) salt solution (TS) to each plate and scraping the surface gently. The resulting slurry was transferred to a sterile test tube and stored at 4°C. Purity was checked on TSA (22°C) by means of the 4 quadrant streak method. Growth on xylose lysine desoxycholate agar (XLD, Oxoid, England) confirmed that Salmonella was the present species (37°C, 24h). Initial cell concentration was determined by plating on XLD and TSA (37°C). Cell suspensions used for inoculation were prepared on the day of use, aiming at around 4 log CFU/sample for inactivation on food simulation media, while this was 5 log CFU/sample for experiments investigating the effect of inoculation method. The solutions were stored at 4°C until further use (max 4h) and concentration N0 was verified by spread plating on TSA at 37°C.
7
Antibiotic Resistance Profiling of Clinical Isolates
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The bacterial strains used in this study are summarized in Table 1 . Escherichia coli and A. baumannii were incubated in LB-Miller medium (10 g/l tryptone (Gibco, Detroit, MI), 5 g/l yeast extract (BD, Sparks, MD), 10 g/l sodium chloride (AppliChem, Darmstadt, Germany)). The remaining four species were incubated in THY broth (36.4 g/l Todd–Hewitt Broth (Oxoid, Basingstoke, UK), 5 g/l yeast extract (BD, Sparks, MD)).
Bacterial strains used in the study and their resistance pattern.
| Species | Strain | Resistance | Source |
|---|---|---|---|
| E. coli | BSU1286* | ESBL | Clinical isolate, Ulm collection |
| S. aureus | ATCC43300 | MRSA | ATCC |
| K. pneumoniae | ATCC700603 | ESBL | ATCC |
| A. baumannii | ATCC19606 | – | ATCC |
| P. aeruginosa | ATCC27853 | – | ATCC |
| E. faecium | DSM17050 | VRE | DSMZ |
| P. aeruginosa | BSU1295** | – | Clinical isolate, Ulm collection |
| P. aeruginosa | BSU1296 | – | Clinical isolate, Ulm collection |
| P. aeruginosa | BSU1297 | – | Clinical isolate, Ulm collection |
| P. aeruginosa | BSU1298 | – | Clinical isolate, Ulm collection |
| S. aureus | DSM 26309 | – | DSMZ |
| S. aureus | ATCC29213 | – | ATCC |
| S. aureus | ATCC13565 | – | ATCC |
| S. aureus | ATCC25923 | – | ATCC |
8
Proteolytic Activity of L. plantarum in WPC
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The medium was prepared by dissolving 50 g of whey protein concentrate (WPC; Marquez Brothers International, Hanford, CA, USA), 5 g of tryptone (Gibco, Paisley, UK), 2.5 g of yeast extract (BD Difco, Detroit, MI, USA), and 1 g of glucose (Daejung Chemicals, Siheung, Korea) in 1 L of distilled water. The medium was sterilized in an autoclave at 95°C for 10 min. To assess proteolytic activity, L. plantarum strains were cultured in MRS and harvested by centrifugation (13,572×g, 4°C, 10 min) and washed twice with PBS. The washed bacteria were resuspended in PBS to approximately 10 Log CFU/mL and 1% (v/v) of the suspension was inoculated into WPC medium (the final concentration of L. plantarum strain was adjusted to 8 Log CFU/mL). The same volume of PBS was used instead of resuspended bacteria as a control. The mixture was incubated at 37°C for 48 h under shaking at low speed. The protein concentration of initial WPC and cultured WPC was measured by the bicinchoninic acid (BCA) assay (Smith et al., 1985) . The ratio of WPC degradation was calculated as follows:
Proteolytic activity(%) = 100 -A 48h / A 0 × 100 (1)
Proteolytic activity(%) = 100 -A 48h / A 0 × 100 (1)
9
Eukaryotic Cell Culture Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
We purchased yeast extract from Thermo Fisher
Scientific, tryptone from Gibco, Sub3 from Bachem, HEK293T/17 cells
from ATCC, Dulbecco’s modified Eagle’s medium (DMEM)
with GlutaMax from Gibco, fetal bovine serum (FBS) from Gibco, polyethylenimine
from Polysciences, and the trypsin-ethylenediamine tetraacetic acid
(EDTA) solution from Gibco. Chemicals used in this work were acquired
from Sigma-Aldrich, Chem Impex, Ambeed, and A2B. Pooled human liver
microsome (1910096) was obtained from Xenotech.
Scientific, tryptone from Gibco, Sub3 from Bachem, HEK293T/17 cells
from ATCC, Dulbecco’s modified Eagle’s medium (DMEM)
with GlutaMax from Gibco, fetal bovine serum (FBS) from Gibco, polyethylenimine
from Polysciences, and the trypsin-ethylenediamine tetraacetic acid
(EDTA) solution from Gibco. Chemicals used in this work were acquired
from Sigma-Aldrich, Chem Impex, Ambeed, and A2B. Pooled human liver
microsome (1910096) was obtained from Xenotech.
10
Bacterial Genetic Manipulation Reagents
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The antibiotics used in this study included ampicillin (Amp), kanamycin (Kan), streptomycin (Sm), and chloramphenicol (Cm). The standard of cembratriene-ol was purchased from Boc Sciences (Shiriey, New York, United States). Yeast extract and tryptone were purchased from Thermo Fisher Scientific Ltd. (Waltham, Massachusetts, USA), glucose, isopropyl-β-d -thiogalactose (IPTG), and L-arabinose were purchased from Sangon Biotech (Shanghai) Co., Ltd.; n-hexane and ethyl acetate were purchased from Shanghai Aladdin Biochemical Technology Co., Ltd.; PCR enzymes (e.g. PrimeSTAR HS DNA polymerase), ligase solution I, and restriction endonucleases needed for molecular biology experiments were purchased from TaKaRa (Dalian, China). The kits for plasmid extraction, agarose gel DNA extraction, and DNA fragment purification were purchased from Axygen Scientific, Inc. (Union, California, USA).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!