The E. coli DH5α bacterial strains were used for cloning and plasmid maintenance. The E. coli strains DH5α and WM335 were used for (−)-α-bisabolol production. Lysogeny broth (LB) medium (containing 10 g/l tryptone, 5 g/l yeast extract, and 10 g/l sodium chloride) was used for all experiments except those with specific indications. MOPS EZ Rich Defined (EZ) medium (Teknova, Hollister, CA, USA) was used per the manufacturer's protocol without ACGU solution. To produce (−)-α-bisabolol, terrific broth containing glycerol (TBG) medium (containing 12 g/l enzymatic casein digest, 24 g/l yeast extract, 9.4 g/l K2HPO4, 2.2 g/l KH2PO4, and 3.5% (w/v) glycerol) was used. SOC medium (containing 20 g/l tryptone, 5 g/l yeast extract, 0.5 g/l sodium chloride, 2.4 g/l magnesium sulfate, 186 mg/l potassium chloride and 4 g/l glucose) was used as a recovery medium after the transformations. l -Rhamnose and antibiotics were purchased from Sigma-Aldrich (St. Louis, MO, USA). Ampicillin was used at a concentration of 100 μg/ml; chloramphenicol at 34 μg/ml; and kanamycin at 25 μg/ml, unless specified otherwise. For the polymerase chain reaction, high fidelity KOD-Plus-Neo polymerase (Toyobo, Osaka, Japan) was used with the standard protocol. All restriction and modification enzymes were purchased from New England BioLabs (NEB, Ipswich, MA, USA).
L rhamnose
Manufactured by Merck Group
Sourced in United States, Germany, China, United Kingdom
L-rhamnose is a monosaccharide that functions as a naturally occurring sugar. It is a component of various plant polysaccharides and can be used in the production of laboratory equipment and reagents.
Automatically generated - may contain errors
Lab products found in correlation
D glucose, by Merck Group (56 mentions)
D mannose, by Merck Group (53 mentions)
D galactose, by Merck Group (53 mentions)
D xylose, by Merck Group (52 mentions)
L arabinose, by Merck Group (36 mentions)
D galacturonic acid, by Merck Group (27 mentions)
D glucuronic acid, by Merck Group (27 mentions)
L fucose, by Merck Group (25 mentions)
D arabinose, by Merck Group (18 mentions)
D fructose, by Merck Group (14 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (8 mentions)
D fucose, by Merck Group (8 mentions)
D ribose, by Merck Group (7 mentions)
Streptomycin, by Thermo Fisher Scientific (6 mentions)
Pullulan standards, by Resonac (5 mentions)
Trifluoroacetic acid, by Merck Group (5 mentions)
Monosaccharide standard, by Merck Group (4 mentions)
Sodium hydroxide (naoh), by Merck Group (4 mentions)
D allose, by Merck Group (4 mentions)
Sodium acetate, by Merck Group (4 mentions)
96 protocols using l rhamnose
1
Production of (-)-alpha-Bisabolol in E. coli
2
CdrA-mediated Aggregation Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The CdrA-mediated aggregation assay was performed as previously described (12 (link)). d -Mannose, l -rhamnose, alginic acid, and sodium alginate from Sigma-Aldrich and uronic acids from Carbosynth LLC were used at 5 mg/mL. Bacterial alginate (Dextra Laboratories Ltd.) was used at 0.5 mg/mL due to poor solubility. Aggregation was visually evaluated and then measured by optical density readings taken at 600 nm. Percent relative aggregation was calculated by taking the difference between the OD600 of the PBAD-cdrAB strain and that of its corresponding vector control strain, dividing by the OD600 of the vector control strain, and multiplying by 100%.
3
Characterization of Ginkgo Seeds
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
G. sinensis and G. microphylla seeds were supplied by Shexian Forestry Bureau in Hebei, China. The samples were collected in December 2012. They were manually separated and kept in a cool and dry place for further use. Moisture content of the whole seed was dried at (105 ± 2 °C) for constant weight (10.0% (w/w)). The standard monosaccharides (l -rhamnose, l -arabinose, d -glucose, d -galactose, d -mannose, and d -xylose) were purchased from Sigma-Aldrich (Saint Quentin Fallavier, France). Dextran standards (DXT3k, DXT25k, DXT160k, DXT760k, and DXT1185k) with an average molecular weight range of 3.7 × 103~1.2 × 106 Da were purchased from TosoHaas (Tokyo, Japan). Absolute ethanol and isopropanol were purchased from Beijing Chemical Works (Beijing, China). Calcium carbonate and concentrated sulfuric acid were all purchased from China National Pharmaceutical Group Corporation (Shanghai, China). Syringe filters were supplied by Tianjin branch billion Lung Experiment Equipment Co. Ltd., (Tianjin, China).
4
Sesame Seed Polysaccharide Extraction
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Sesame seeds (Sesamum indicum L.) were obtained from the Sesame Research Center, Henan Academy of Agricultural Sciences. After sieving to remove impurities, the seeds were dried for 24 h at 40°C and then kept in a sealed container at 4°C until decortication. Chemicals required for the assays, including dextran standards and standard monosaccharides (D-glucose, D-galactose, D-mannose,D-xylose,L-rhamnose, L-arabinose, D-galacturonic acid, D-glucuronic acid) were purchased from Sigma Chemical Co. (St. Louis, MO, United States). DEAE Cellulose-52 was obtained from Solarbio Science and Technology Ltd. (Beijing, China). All other chemical reagents were acquired from Tianjin Chemical Reagents Co. Ltd. (Tianjin, China) of analytical grade.
5
Isolation and Characterization of Dextran from Agaricus bisporus
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fruiting bodies of A. bisporus (Yingxiu 1#) were collected from Zhejiang Agriculture Research Institute in Zhejiang province, China. T-series dextrans were purchased from Wuhan Putus Macromolecular Sci. & Tech. Co. Ltd., (Wuhan, China). Monosaccharide standards (d -galactose, d -arabinose, l -fucose, l -rhamnose, d -manonose, d -xylose, d -glucose and erythrose), trifluoroacetic acid (TFA) and dimethyl sulfoxide (DMSO) were purchased from Sigma (St. Louis, MO, USA). All other chemical reagents were of grade AR from Shanghai Chemical Co. (Shanghai, China). DEAE-Sepharose Fast Flow and Sephacryl S-300 High Resolution were purchased from Amersham Pharmacia Biotech (Uppsala, Sweden). TSKgel G4000PWXL column was purchased from Shanghai Biological Technology Co., Ltd. (Shanghai, China). Ultrahydrogel TM 120 and 1000 (78 × 300 mm) columns were purchased from Waters Co., Milford, MA, USA. HPLC was carried out on a waters 1525 HPLC system (1525 HPLC pump, 2414 refractive index detector; Waters Co., Milford, MA, USA).
6
Purification of His-tagged Protein from E. coli
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A single colony of E. coli harboring pRham N-His Kan::LysKB317 (Table 1 ) was isolated and inoculated into 5 mL LB media with Kan at 37°C and 200 rpm shaking overnight. This was used to inoculate fresh 25 mL LB with Kan at a ratio of 1:100, which was then incubated at 37°C and 200 rpm until the OD600 nm reached 0.6 (Lu et al., 2020 (link)). Culture was induced with 0.2% (w/v) L-rhamnose (Sigma-Aldrich Inc.) for at least 16–18 h. Cells were centrifuged at 8,000 x g for 15 min at 4°C, and the spent medium was decanted. Cell pellets were then lysed using B-Per (Invitrogen) with DNase I, RNase I, and lysozyme (10 U/mL; ThermoFisher) and incubated at room temperature for 20 min. The whole cell lysate was centrifuged at 15,000 x g for 5 min and the soluble supernatant was pipetted into a His-Spin column (ZymoResearch) following the manufacturer’s protocol for Ni-NTA protein purification. The concentration of purified protein was quantified using the Qubit Protein Assay Kit (ThermoFisher) and subsequently used as a positive control for SDS-PAGE and Western blot analysis.
7
Fluorescence Imaging of Fungal Strains
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Strains IPO323_Picl1G_mChSso1, IPO323_Pgal7G_mChSso1, IPO323_Pex1AG_mChSso1 and IPO323_PlaraBG_mChSso1 were grown in minimal medium without carbon source or supplemented with 2% (w v−1) glucose, galactose, fructose, sucrose, xylose, L-rhamnose and L-arabinose (Sigma–Aldrich) for 2 days at 18 °C, 200 rpm. Aliquots of these cultures were investigated using laser-supported epifluorescence as described previously (Schuster et al., 2015a (link)). 14-bit images were acquired at various laser intensities (percent output power: Pex1A, 30%; PlaraB, 100%; Pgal7, 40%; Picl1, 20%) and 300 ms exposure time. Signal intensities in these images were measured using the software package Metamorph 7.8x.
8
Anticoagulant Potential of Seaweed M. angicava
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Seaweed M. angicava was collected from the coast of Qingdao, China on April 2013, which is in growth mature period of the seaweed. The raw material was thoroughly washed with tap water, air-dried and stored at room temperature in a dry environment. l -rhamnose, l -arabinose, d -xylose, l -fucose, d -mannose, d -galactose, d -glucose, d -glucuronic acid, d -galacturonic acid, d -glucosamine and heparin were from Sigma-Aldrich Chemical Co. (St. Louis, MO, USA). Pullulan standards (Mw: 21.1, 47.1, 107, 200, 344, and 708 kDa) were from Showa Denko K.K. (Tokyo, Japan). APTT assay reagent (ellagic acid + bovine phospholipids reagent), TT assay reagent (bovine thrombin) and PT assay reagent (rabbit thromboplastin) were from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). PAI-1 and D-dimer kits were from Simens Healthcare Diagnostics Products (Marburg, Germany). FDP kit was from BIOLINKS CO., LTD. (Tokyo, Japan).
9
FDG-based Bacterial Protein Visualization
10
Preparation of Antibiotic Stock Solutions
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
C109 was synthesized as previously described (12 (link)). Antibiotic working stock solutions were prepared as follows: C109, 10 mg/ml in dimethyl sulfoxide (DMSO); trimethoprim, 50 mg/ml in DMSO (Sigma); doxycycline hyclate, 25 mg/ml in H2O (Sigma); chloramphenicol, 20 mg/ml in ethanol (EtOH; Sigma), meropenem, 10 mg/ml in DMSO (Sigma); tobramycin sulfate, 10 mg/ml in H2O (Alfa Aesar); ceftazidime, 10 mg/ml in 0.1 M NaOH (Sigma); ciprofloxacin, 10 mg/ml in 0.1 M HCl (Sigma); novobiocin sodium, 10 mg/ml in H2O (Sigma); and piperacillin sodium salt, 10 mg/ml in H2O (Sigma). IPTG and L-rhamnose were obtained from Sigma.
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!