The candidate bacterial endophyte 3F11 along with a negative control endophyte 3F7 (Supplementary Table S1 ) were grown overnight in LB medium at 37 °C with shaking at 250 rpm. Ten microliters from overnight cultures (adjusted to OD595 = 1.0) were spotted onto NBRIP agar as described above, but supplemented with 15 g Bacto agar and 0.4 g bromocresol purple (Sigma, B5880) per liter. bromocresol purple was used as a pH indicator (changes from purple to yellow at pH 6.8 to 5.2). Glucose (filter sterilized) and bromocresol purple were added after autoclaving and cooling of the medium to 55 °C. The plates were incubated at 28 °C and observed for changes in colour after 5 h and 24 h. There were 4 replicate colonies in a Petri dish in the first trial and 4 replicate colonies in the second trial.
Bromocresol purple
Manufactured by Merck Group
Sourced in United States, Germany
Bromocresol purple is a pH indicator dye commonly used in analytical chemistry and biochemistry applications. It exhibits different colors within a specific pH range, allowing it to be used for pH measurement and titration. Bromocresol purple changes color from yellow to purple as the pH increases from approximately 5.2 to 6.8. This characteristic makes it a useful tool for various laboratory procedures, but its specific applications should be determined by the user's needs and requirements.
Automatically generated - may contain errors
Lab products found in correlation
Bromocresol green, by Merck Group (3 mentions)
Methyl red, by Merck Group (3 mentions)
Cresol red, by Merck Group (2 mentions)
Neutral red, by Merck Group (2 mentions)
Bromothymol blue, by Merck Group (2 mentions)
Thymol blue, by Merck Group (2 mentions)
L arginine, by Merck Group (2 mentions)
Histidine, by Merck Group (1 mentions)
Ornithine, by Merck Group (1 mentions)
Lysine, by Merck Group (1 mentions)
Tyrosine, by Merck Group (1 mentions)
Glycerol, by Merck Group (1 mentions)
P coumaric acid, by Merck Group (1 mentions)
Alizarin, by Merck Group (1 mentions)
Polyvinylidene fluoride pvdf membrane, by Merck Group (1 mentions)
Methyl red sodium salt, by Merck Group (1 mentions)
Citric acid, by Scharlab (1 mentions)
Formic acid, by Thermo Fisher Scientific (1 mentions)
Glycerol, by Scharlab (1 mentions)
Methyl orange, by Merck Group (1 mentions)
26 protocols using bromocresol purple
1
Bacterial Endophyte Phosphate Solubilization Assay
2
Screening Non-Conventional Yeasts for Biogenic Amines
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To confirm that the non-conventional yeast strains shortlisted here (Table 1 ) are safe to use, we checked that none produced any biogenic amines (BAs). Production of BAs was determined using an adapted version of the method explained by Joosten and Northholt [28 (link), 29 (link)]. Briefly, yeast strains (106 cells per ml) were inoculated onto YPD agar plates supplemented with bromocresol purple (Sigma Aldrich) 0.006% and an amino acid mix with a total mass concentration of 1% (MP Biochemicals, LLC). The added amino acids are tyrosine, histidine, phenylalanine, leucine, tryptophan, arginine and lysine at equal ratios. Subsequently, the plates were incubated at 30°C for 7 days and the growth and changes in the color of the medium was monitored daily to test for the presence of BAs. In strains with no BA production, the growth area was surrounded by a yellow halo caused by glucose fermentation, followed by a pH reduction that causes the medium to turn purple after a period that depends on the growth rate of the strain. By contrast, when BAs are produced, amino acid decarboxylation resulted in a purple halo from the very beginning, which grew bigger and darker as a function of time.
3
Temperature-dependent Growth Profiling
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The isolates were evaluated for their ability to grow at 35, 37, 40, and 45 °C in MRS broth (pH 6.8) containing bromocresol purple (Sigma Aldrich, 0.16 g/L) as a pH indicator. The inoculated broths were incubated for 48 h (35–45 °C). At the end of the incubation period, a color change from purple to yellow due to broth acidification was indicative of bacterial growth [28 (link)].
4
Rice Rhizosphere Acidification Assay
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Rice rhizosphere acidification assays were performed as described previously (Pacheco-Villalobos et al., 2016 (link)). For assessment of rhizosphere acidification by different plant lines, we transferred healthy 7-day-old rice seedlings to plates containing half-strength Murashige and Skoog agar supplemented with 0.15 mM bromocresol purple (Sigma-Aldrich; sensitivity range, pH 5.2–6.8). The plates were incubated in an artificial climate chamber with a 16-h-light (30°C)/8-h-dark (22°C) photoperiod and 60%–70% relative humidity for 24 h and then scanned.
5
Biogenic Amine Production Assay
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L. plantarum K10 grown in MRS broth at 37°C for 18 h were
cultured in a special medium according to Chang and Chang’s method (2012)
for 24 h at 37°C. LB agar (pH 5.0; Difco) containing 0.25% glycerol,
0.006% bromocresol purple, and 0.1% precursor amino acid (tyrosine, histidine,
ornithine, and lysine, respectively; Sigma) were used. The strain was washed
three times with distilled water, and 10 µL of sample was loaded onto
sterile filter paper discs (8 mm diameter, Advantec, Japan). Biogenic amine
production was determined by the color change of the pH indicator bromocresol
purple in the medium.
cultured in a special medium according to Chang and Chang’s method (2012)
for 24 h at 37°C. LB agar (pH 5.0; Difco) containing 0.25% glycerol,
0.006% bromocresol purple, and 0.1% precursor amino acid (tyrosine, histidine,
ornithine, and lysine, respectively; Sigma) were used. The strain was washed
three times with distilled water, and 10 µL of sample was loaded onto
sterile filter paper discs (8 mm diameter, Advantec, Japan). Biogenic amine
production was determined by the color change of the pH indicator bromocresol
purple in the medium.
6
Screening Lactobacillus for Biogenic Amine Production
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The ability of Lactobacillus strains to produce BA by decarboxylation of amino acids was tested on a media designed by Yousif et al. (2005 (link)) which contained the precursor amino acids including L‐histidine monohydrochloride, tyrosine di‐sodium salt, L‐ornithine monohydrochloride, and L‐lysine monohydrochloride (Acros & Bio basic). First, Lactobacillus spp. were subcultured twice in MRS broth containing 0.1% of each precursor amino acid and 0.005% pyridoxal‐5‐phosphate. After that, strains were spotted on the MRS agars with and without amino acids which containing 0.06% bromocresol purple (Sigma). After 2–5 days incubation, purple color obtained in surrounding colonies was considered as a positive (Yousif et al., 2005 (link)).
7
Screening Yeast Decarboxylase Activity
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YPD agar supplemented with 0.145% (p/v) p-coumaric acid (Sigma-Aldrich) and 0.01% (w/v) bromocresol purple (Sigma-Aldrich) was used to evaluate HCDC activity [13 (link)]. A single colony was spread onto the plates and was incubated at 25 °C for 5–7 days. At the time of inoculation, the colour of the medium was purple. The activity of isolates resulted in an acidification of the medium, turning it yellow. If the decarboxylase activity was positive, the agar surrounding the colonies turned purple.
8
Colorimetric pH Indicator Development
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BDA was purchased from Wan-do Kumil-Suhyup (Wan-do, Jeollanam-do, Republic of Korea). Nine pH indicators (thymol blue, bromocresol green, cresol red, bromocresol purple, neutral red, bromoxylenol blue, alizarin, methyl red, and metanil yellow) were purchased from Sigma Aldrich Chemical Co., Ltd. (St. Louis, MO, USA). Ethanol (99.9%) was obtained from Duksan Pure Chemicals (Ansan-si, Gyeonggi-do, Korea). Polyvinylidene fluoride (PVDF) membranes were sourced from Merck KGaA (Darmstadt, Hesse, Germany). Sodium alginate, glycerol, calcium chloride, magnesium oxide, glacial acetic acid, chloroform, sodium thiosulfate, potassium iodide, trichloroacetic acid (TCA), butylated hydroxytoluene (BHT), and thiobarbituric acid (TBA) were purchased from Duksan Science (Ansan, Gyeonggi, Republic of Korea).
9
pH-Sensitive Textile Sensors Development
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The cotton and polyester jersey knits were provided by A. Ferreira & Filhos. One knit was composed of 100% polyester with a thickness of 0.68 mm and a mass per unit area of 207 g/m2 and the other of 100% cotton with a thickness of 0.89 mm and a mass per unit area of 187 g/m2. The pH indicators used for the development of the sensors were methyl red (C15H15N3O2), methyl red sodium salt (C15H15N3O2Na), methyl orange (C14H14N3NaO3S), bromocresol purple (C21H16Br2O5S), bromothymol blue (C27H28Br2O5S) and were purchased from Sigma-Aldrich (St. Louis, Mo., EUA). The chemicals used in the evaluation of samples sensitivity were the following: acetic acid acquired from Chem-lab (Zedelgem, Belgium); sodium hydroxide purchased from Normax (Marinha Grande, Portugal); formic acid obtained from Fisher Scientific (Hampton, NH., EUA); ammonia purchased from VWR Chemicals (Radnor, PA, EUA); and citric acid acquired from Scharlau (Barcelona, Spain). The polyurethane polymeric base employed in the preparation of pH-sensitive formulations before application on fibrous substrates, Edolan SN and Thickener A02 were purchased from ADI Center Portugal, S.A (Santo Tirso, Portugal). Glycerol was obtained from Scharlau. The filter paper, used as substrate in one part of this work, was purchased from Normax.
10
Polysaccharide Utilization by Xylose-Fermenting LAB
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The selected strains of xylose utilizing LAB from Eri silkworm midgut were investigated for their abilities to utilize the selected polysaccharides as the sole carbon source. Briefly, individual LAB isolate was spiked on MRS agar containing 0.5% (w/v) of single carbon source of individual polysaccharides including starch (Fisher Scientific, Loughborough, Leicestershire, UK), pectin (Fisher Scientific, Loughborough, Leicestershire, UK), beechwood xylan (Megazyme International, Bray, Co. Wicklow, Ireland), locust bean gum (Sigma Aldrich, St. Louis, MO, USA) or carboxymethyl cellulose (Sigma Aldrich, St. Louis, MO, USA) supplemented with bromocresol purple (0.04%, w/v). After incubation at 37 °C for 24 h, the bacterial growth and the yellow halo-formed surrounding colonies were observed. Further duplicated experiment was also carried out, but the bromocresol purple was replaced with trypan blue (0.01%, w/v) for detection of extracellular activities of amylase, pectinase, xylanase, β-mannanase, and cellulase, respectively. The clear zone formed surrounding colonies were observed after incubation at 37 °C for 24 h.
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