Parental SGC-7901, SGC-7901/B1 and subline SGC-7901/B2 cells were suspended in 0.3% agar (Amresco, America), respectively. Cells (1000 cells/well) were seeded in a six-well plate containing an underlayer of 0.6% agar and incubated at 37 °C in a humidified atmosphere of 5% CO2. After two weeks of incubation, colonies containing more than 50 cells were counted. The colony formation efficiency was measured as the average percentage of three wells.
Agar
Manufactured by Avantor
Sourced in United States, Germany
Agar is a gelatinous substance derived from red algae. It is commonly used as a solidifying agent in microbiological culture media, allowing the growth and isolation of bacteria and other microorganisms.
Automatically generated - may contain errors
Lab products found in correlation
Glucose, by Merck Group (4 mentions)
Yeast extract, by Avantor (3 mentions)
Congo red, by Carl Roth (2 mentions)
Sodium hydroxide (naoh), by Merck Group (2 mentions)
Alginate, by Merck Group (1 mentions)
Trizol reagent, by Merck Group (1 mentions)
Sterile glycerol, by ICN Biomedicals (1 mentions)
Tween 20 solution, by Merck Group (1 mentions)
Yeast extract, by BD (1 mentions)
Czapek dox broth, by BD (1 mentions)
Propylene oxide, by Merck Group (1 mentions)
Nystatin, by Merck Group (1 mentions)
Nalidixic acid, by Merck Group (1 mentions)
Cycloheximide (chx), by Merck Group (1 mentions)
Tween 80, by Merck Group (1 mentions)
Mgso4.7h2o, by Avantor (1 mentions)
Nano3, by Avantor (1 mentions)
Potassium chloride (kcl), by Avantor (1 mentions)
K2hpo4, by Avantor (1 mentions)
Potassium iodide, by Avantor (1 mentions)
30 protocols using agar
1
Soft Agar Colony Formation Assay
2
Soft Agar Colony Formation Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Transfected cells were propagated on soft agar. A base layer (1.5 mL) of agar (Amresco, Solon, OH) (0.6% agar in RPMI-1640 with 10% FBS) was allowed to solidify in a six-well flat-bottomed plate before the addition of 2 mL of a cell suspension containing 2000 cells in 0.3% agar in RPMI-1640 with 10% FBS. The colonies were allowed to grow for 28 days at 37°C under 5% CO2 and visualized with an inverted microscope (Olympus IX71, Olympus, Tokyo, Japan).
3
Comparative Growth of Z. galactanivorans on Diverse Polysaccharides
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The substrates tested for growth were glucose (Merck), alginate (Sigma), laminarin (Degussa), ι-carrageenan (Sanofi BioIndustries), κ-carrageenan (extracted from Eucheuma cottonii, gift from Goëmar), agar (Amresco), or porphyran (extracted from Porphyra umbilicalis as in Correc et al., 2011 (link)). 1H NMR analysis of the self-extracted porphyran and κ-carrageenan confirmed the purity and the motif composition of the polysaccharides (Correc et al., 2011 (link); Préchoux et al., 2016 (link)). The type strain Z. galactanivorans DsijT (Barbeyron et al., 2001 (link)) was routinely grown from glycerol stocks in Zobell medium 2216E (Zobell, 1941 ) at 20°C. Cells were pelleted (5 min, 4,000 g) and resuspended in saline solution to remove traces of organic substrates. This suspension was inoculated (1/50 ratio) in triplicate flasks containing marine mineral medium (Thomas et al., 2011a (link)) amended with 2 g.L−1 of one substrate as sole carbon source. Cultures were incubated at 20°C, 180 rpm. After 48 h, aliquots (5 ml) were centrifuged (10 min, 4,000 g) and cell pellets were resuspended in 200 μl Trizol reagent (Sigma), frozen in liquid nitrogen and stored at −80°C until RNA extraction.
4
Cryogenic Storage of Fungal Spores
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Spores were collected after 2 weeks of incubation at 24°C on Czapek yeast extract medium (CZY: 35 g/L czapek dox broth, Difco Laboratories, United States; 2 g/L yeast extract, Difco Laboratories, United States; 15 g/L agar, Amresco, United States; pH 6.5) scraping the surface of the colonies with a sterile loop and 5 mL of 10% sterile glycerol (ICN Biomedicals, United States) + 0.01% Tween20 solution (Sigma-Aldrich, United States). The concentration was determined using a hemocytometer and adjusted to 107 spores/mL. Small aliquots were then stored at −20°C.
5
Streptomyces Colonization in Wheat Roots
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Control plants and plants inoculated only with Streptomyces (no-Fusarium inoculation) from the previous test were harvested 20 days after transplant and washed in sterile water to remove the excess soil. Shoot length and dried weight of wheat plants were assessed for each treatment.
For inner root tissue analysis, 10 seedlings for each treatment were selected and cut at the base. The roots were washed and surface sterilized with propylene oxide (Sigma-Aldrich, United States) for 1 h (Sardi et al., 1992 (link)). Subsequently, 10 or 15 root pieces were cut in sterile conditions and placed in water agar medium (WA) containing 15 g/L agar (Amresco, Italia), 25 mg/L nalidixic acid (Sigma-Aldrich, United States), 50 mg/L nystatin (Sigma-Aldrich, United States), and 50 mg/L cycloheximide (Sigma-Aldrich, United States). Plates were incubated for 7 days at 24°C. Growth of Streptomyces spp. colonies on the plate was visually observed using a microscope. Morphological examination was carried out to confirm the re-isolation. Roots not inoculated with Streptomyces strains were used as negative control and subjected to the same procedure to check the presence of Streptomyces spp.
For inner root tissue analysis, 10 seedlings for each treatment were selected and cut at the base. The roots were washed and surface sterilized with propylene oxide (Sigma-Aldrich, United States) for 1 h (Sardi et al., 1992 (link)). Subsequently, 10 or 15 root pieces were cut in sterile conditions and placed in water agar medium (WA) containing 15 g/L agar (Amresco, Italia), 25 mg/L nalidixic acid (Sigma-Aldrich, United States), 50 mg/L nystatin (Sigma-Aldrich, United States), and 50 mg/L cycloheximide (Sigma-Aldrich, United States). Plates were incubated for 7 days at 24°C. Growth of Streptomyces spp. colonies on the plate was visually observed using a microscope. Morphological examination was carried out to confirm the re-isolation. Roots not inoculated with Streptomyces strains were used as negative control and subjected to the same procedure to check the presence of Streptomyces spp.
6
Fungal Diversity Isolation from Dust
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The settled dust samples, c. 10 mg, were seeded on plates containing a malt extract medium (15 g malt extract from Sharlab, Barcelona, Spain and 12 g of agar from Amresco, Dallas, USA, in 500 mL of H2O) or a tryptic soy extract agar (Sharlab, Barcelona, Spain, 20 g L−1 in 500 mL) medium without antibiotics or fungicides and were sealed at the site of sampling with gas-permeable adhesive tape. Colonies were inspected and counted after one, two, and three weeks of culturing at 23 ± 2 °C. After three weeks of incubation, the colonies on the primary isolation plates (not yet single-spored) were numbered and screened for toxicity.
The ability to grow at 37 °C was tested on malt extract agar plates sealed with gas-permeable tape, which were then incubated for five days. The strains Trichoderma longibrachiatum THG and T. atroviride H1/226 were used as positive and negative controls, respectively [17 (link)]
The ability to grow at 37 °C was tested on malt extract agar plates sealed with gas-permeable tape, which were then incubated for five days. The strains Trichoderma longibrachiatum THG and T. atroviride H1/226 were used as positive and negative controls, respectively [17 (link)]
7
Porcine Semen and Cell Culture Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Boar semen, 27 × 106 sperms per ml in MRA extender, was purchased from Figen Ltd., Tuomikylä, Finland. The porcine kidney (PK‐15), murine neuroblastoma (MNA) and feline lung (FFL) cells retrieved from EVIRA (Echard 1974 ; Andersson et al. 2009 ) were grown in a tissue culture facility as described in detail by Ajao et al. (2015 ). Malt extract agar media contained 15 g malt extract (Sharlab, Barcelona, Spain) and 12 g agar (Amresco, Dallas, USA) in 500 ml of H2O. Tween 80 was from Sigma Aldrich, St Louis, Missouri, USA.
The UV illuminator was from UVA Finland Ltd., Kauniainen. The toxic fungal droplets were photographed using a Dino‐Lite microscopic loupe (Taiwan), magnification 200 ×, connected by USB to a laptop computer.
The UV illuminator was from UVA Finland Ltd., Kauniainen. The toxic fungal droplets were photographed using a Dino‐Lite microscopic loupe (Taiwan), magnification 200 ×, connected by USB to a laptop computer.
8
Screening Cellulase-Producing Bacteria
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cellulase-producing bacteria were screened on carboxymethyl cellulose (CMC) agar plates containing: low-viscosity carboxymethyl cellulose sodium salt 2 g/L (Glentham Life Sciences, Corsham, UK), NaNO3 2 g/L, K2HPO4 1 g/L, MgSO4.7H2O 0.5 g/L, KCl 0.5 g/L (all from Chempur, Piekary Slaskie), peptone 0.2 g/L (BTL) and agar 2 g/L (VWR; Avantor, Radnor, PA, USA) (28 (link)). Strains were activated during 24 h of cultivation at 50 °C in nutrient broth. Then, the microorganisms were plated onto CMC agar and incubated at 50 °C for 2 days. Resulting colonies were picked up and transferred onto a new CMC agar to ensure that bacterial growth was a result of CMC degradation. After 3 days of incubation at 50 °C, the zones surrounding the colonies were visualised by Congo red (Carl-Roth GmbH & Co. KG, Karlsruhe, Germany) staining (29 (link)). To compare the capability of CMC degradation, hydrolysis capacity was calculated. The hydrolysis capacity is defined as the ratio of the diameter of clearing zone around the colony and the colony diameter (30 (link)).
9
Synthesis and Analysis of Raw-Re Nanostructures
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All of the reagents were of an analytical grade or better. The de-ionized water was used in all the experiments. The raw-Re nanostructures were obtained from the precursor solution of ammonium perrhenate(vii) (NH4ReO4), which contains 2000 mg L−1 of Re(vii ) ions. The reagents for the determination of the selected RNS and ROS were purchased from Hanna Instruments (Salaj, Romania) and Sigma-Aldrich (Steinheim, Germany), respectively. The reagents used for ROS determination were: (i) potassium iodide, (ii) starch, and (iii) agar, and they were purchased from Avantor Performance Materials (Gliwice, Poland), Sigma-Aldrich (Steinheim, Germany), and BTL (Lodz, Poland), respectively. The 4-NP and NaBH4 (MERCK, Branch Poland) used for the catalytic hydrogenation were applied as received.
10
Cultivation of Bacterial Strains
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Bacterial strains used in this study are listed in Table 2 . The following strains were cultivated in Brain heart infusion broth (Thermo Scientific Oxoid ): enterococcal strains (37 °C, without agitation), staphylococcal strains (37 °C, 220 rpm), and E. coli (37 °C, 220 rpm). L. plantarum and L. lactis were cultivated without shaking in DeMan, Rogosa and Sharp (MRS) broth (Thermo Scientific Oxoid ) at 37 °C and M17 broth (Thermo Scientific Oxoid ) supplemented with 0.5% glucose at 30 °C, respectively. agar plates were prepared by supplementing the appropriate broth with 1.5% (w/v) agar (VWR chemicals). Erythromycin was added to a final concentration of 200 μg/ml for E. coli and 10 μg/ml for L. plantarum when appropriate.
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!