Du730

Manufactured by Beckman Coulter

Sourced in United States, Germany, United Kingdom

The DU730 is a UV/Vis spectrophotometer manufactured by Beckman Coulter. It is designed to measure the absorbance or transmittance of light passing through a sample. The instrument covers a wavelength range of 190 to 1100 nanometers and can be used for a variety of analytical applications in laboratory settings.

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DU730 Life Sciences Model DU730 DU730 Life Science UV/Vis DU730 spectrophotometer

166 protocols using du730

Quantitative Analysis of Total Carbohydrates and Starch

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Total carbohydrate in cells was determined according to the colorimetric method using a UV/Vis spectrometer (DU 730, Beckman Coulter, USA), as described by Kang et al. [36 (link)]. Freeze-dried biomass was re-suspended in 10 mL of DW, and then 1 mL of samples was mixed with 1 mL of 5% (w/v) phenol solution. Then, 5 mL of highly concentrated sulfuric acid (95–98%) was added and incubated for 30 min at ambient temperature. After slight vortexing, optical density was measured at 470 nm using a UV/Vis spectrometer (DU 730, Beckman Coulter, USA). The amount of total carbohydrate was calculated from a standard which is made by glucose.
Starch was assayed by an enzymatic method using degradation of the starch to glucose with α-amylase and amyloglucosidase. Ground lyophilized cells were mixed with 10 mL of 80% (v/v) ethanol, and incubated at 80 °C for 60 min to remove interfering materials. Quantification of the starch percentage was conducted using a total starch assay kit (K-TSTA-100A, Megazyme, USA) according to the manufacturer’s instructions.

Oh S.H., Chang Y.K, & Lee J.H. (2019). Identification of significant proxy variable for the physiological status affecting salt stress-induced lipid accumulation in Chlorella sorokiniana HS1. Biotechnology for Biofuels, 12, 242.

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Terpenoid and Lipid Content Analysis

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Terpenoid content in sample extract was tested using linalool at different concentrations as standard reference to obtain the calibration curve. 1 mL of each sample extract was prepared in respective vials and mixed with 1.5 mL of chloroform. After gently mixing, 3 drops of sulphuric acid (95%) were slowly added into all mixtures. Finally, the absorbance readings of all samples and standard solutions were detected using UV–VIS Spectrophotometer (Beckman Coulter/DU 730) at a fixed wavelength of 538 nm against blank sample (Ramnath et al. 2015 ).
The assay of lipid content was carried out by preparing 0.75 g of vanillin reagent diluted with 125 mL of distilled water as a stock solution. Pure olive oil was prepared at different concentrations (0.2–1 mL) as a standard reference to perform the calibration curve. Next, 1 mL of each sample extract and standard solution were added with 1.5 mL H₂SO₄ (95%) before incubated in a water bath (Bath WB Water Bath 18/30/45 Litre 100 °C ± 0, 1 °C) at 60 °C for 10 min. After cooling, 2.4 mL of vanillin reagent was added into each mixture and incubated at room temperature for 40 min. The absorbance was measured through UV–VIS Spectrophotometer (Beckman Coulter/DU 730) at a fixed wavelength of 490 nm (Abdullah et al. 2020 (link)).

Salleh S.N., Hanapiah N.A., Johari W.L., Ahmad H, & Osman N.H. (2021). Analysis of bioactive compounds and chemical composition of Malaysian stingless bee propolis water extracts. Saudi Journal of Biological Sciences, 28(12), 6705-6710.

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Rapamycin Nanoparticle Characterization and Release

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RAPNPs were first frozen at -80 °C and then freeze-dried with a Labconco Free Zone lyophilizer. Then, the RAPNP lyophilized powder was dissolved in DMSO, and the absorbance was measured with a UV/Vis spectrophotometer (DU730, Beckman Coulter) at 280 nm. According to the preestablished standard curve of RAP in DMSO, the drug loading efficiency (LE) and drug encapsulation efficiency (EE) were calculated as follows:
where M_RAP is the mass of RAP loaded in the NPs, M_PLGA is the mass of polymer in the formulation and M_added is the mass of RAP added.
The drug (RAP) release from RAPNPs and MM/RAPNPs was studied separately using a dialysis method. Briefly, RAPNP and MM/RAPNP solutions (2 mg/mL, 1 mL each) were added to disposable dialysis bags (MWCO: 3500 Da, Thermo Scientific). The dialysis bags were then immersed in 10 mL of phosphate-buffered saline (PBS) solution (release medium, pH 7.4) at 37 °C. Three independent replicates were used for each sample. One milliliter of release medium was collected for analysis at different time intervals and replaced with an equivalent volume of fresh PBS at 37 °C. The cumulative amount of RAP released was quantified by a UV/Vis spectrophotometer (DU730, Beckman Coulter) at 280 nm.

Wang Y., Zhang K., Li T., Maruf A., Qin X., Luo L., Zhong Y., Qiu J., McGinty S., Pontrelli G., Liao X., Wu W, & Wang G. (2021). Macrophage membrane functionalized biomimetic nanoparticles for targeted anti-atherosclerosis applications. Theranostics, 11(1), 164-180.

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Measuring Drug Encapsulation and Release

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The drug loading efficiency (DLE) and drug encapsulation efficiency (DEE) were measured using UV/Vis Spectrophotometer (DU730, Beckman Coulter) at a 292-nm wavelength and calculated according to the standard curve and equations (1) and (2) to get the final result.

DLE = \frac{Mass of RAP}{Mass of PCD + Mass of RAP} \times 100 %

DEE = \frac{Mass of RAP}{Mass of the added RAP} \times 100 %

Drug release from RNPs and MM/RNPs were characterized using a dialysis method. Briefly, 1 ml of NPs solutions (500 µg/ml) was put into a dialysis bag (MWCO 3500 Da, Solarbio) and incubated in 10 ml of release medium (PBS: DMSO = 8:2, v/v) with or without H₂O₂, at predetermined time points, 1 ml of fresh medium was replenished after 1 ml of the original release medium was withdrawn for UV/Vis analysis. The cumulative amount of released RAP was measured using UV/Vis Spectrophotometer (DU730, Beckman Coulter) at a 280 nm wavelength and calculated based on the standard curve and equation (3).

Cumulative drug release = \frac{M_{t}}{M_{0}} \times 100 %

where M_t and M₀ represent the amount of drug released at time t and the initial amount of drug in the NPs, respectively.

Tang D., Wang Y., Wijaya A., Liu B., Maruf A., Wang J., Xu J., Liao X., Wu W, & Wang G. (2021). ROS-responsive biomimetic nanoparticles for potential application in targeted anti-atherosclerosis. Regenerative Biomaterials, 8(4), rbab033.

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Measuring Film Transparency using Spectrophotometry

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The opacity of each film was measured to determine the transparency of each film. A spectrophotometer (BECKMAN COULTER/DU 730, Beckman Coulter, Brea, CA, USA) was used to measure the transparency according to Equation (5).

Opacity = \frac{Abs 600}{x}

where x represents the thickness (in mm) of the film, and Abs600 is the absorbance of light measured at 600 nm.

Mohammed A.A., Hasan Z., Omran A.A., Elfaghi A.M., Khattak M.A., Ilyas R.A, & Sapuan S.M. (2022). Effect of Various Plasticizers in Different Concentrations on Physical, Thermal, Mechanical, and Structural Properties of Wheat Starch-Based Films. Polymers, 15(1), 63.

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ESBL Detection and Plasmid Isolation

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K. pneumoniae-produced ESBLs were detected using a commercial ESBL Assay Kit (Hangzhou Binhe Microorganism Reagent Co., Ltd., Hangzhou, China) , according to the manufacturer protocols. The variable temperature-sodium dodecyl sulfate elimination method was used to isolate the plasmids from ESBLs; bacterial DNA was extracted using a DNA extraction kit (Tiangen Biotech Co., Ltd., Beijing, China) , and the plasmid DNA extracted using a plasmid extraction kit (Tiangen Biotech Co., Ltd.). The concentration and purity of DNA was determined using a nucleic acid detector of BeckMan Coulter DU730 (Beckman Coulter, Inc., Brea, CA, USA); the DNA was stored at -20°C until further use.

Huang S.Y., Pan K.Y., Liu X.Q., Xie X.Y., Dai X.L., Chen B.J., Wu X.Q, & Li H.Y. (2015). Analysis of the drug-resistant characteristics of Klebsiella pneumoniae isolated from the respiratory tract and CTX-M ESBL genes. Genetics and molecular research : GMR, 14(4).

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Antioxidant Enzyme Activity Assay

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The 8-day-old seedlings grown on 0%, 0.5%, or 2% sucrose 1/2 MS medium were harvested and total proteins were extracted at 4 °C. Superoxide dismutase (SOD) and catalase (CAT) activities were measured with SOD and CAT assay kits (Comin Biotechnology Co., Ltd., Suzhou, China) as described [30 (link)]. A spectrophotometer (DU730, Beckman Coulter, Pasadena, CA, USA) was used to measure the absorbances at 560 nm and 240 nm for SOD and CAT, respectively.

Yuan Y., Xu D., Xiang D., Jiang L, & Hu H. (2022). Serine Hydroxymethyltransferase 1 Is Essential for Primary-Root Growth at Low-Sucrose Conditions. International Journal of Molecular Sciences, 23(9), 4540.

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Analytical Methods for Biofuel Production

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Cell density (OD₆₀₀) was determined by spectrophotometer (DU730, Beckman Coulter). Assays of products (butanol, acetone and ethanol) were performed as previously described [32 (link)]. In brief, samples were taken at appropriate time intervals and then centrifuged at 7, 000 g for 20 min at 4 °C. The products in supernatant were then analyzed using a gas chromatograph (7890 A, Agilent, Wilmington, DE, USA) equipped with a flame ionization detector (Agilent) and a capillary column (EC-Wax; Alltech), details were described previously [32 (link)].
Glucose and formate were determined by HPLC (model 1200, Agilent, USA) equipped with an Aminex HPX-87H ion-exclusion column (Bio-Rad) and a differential refractive index detector (RID). 5 mM of H₂SO₄ was used as mobile phase at 0.6 mL/min flow rate. The column temperature was set at 30 °C.

Zhao R., Dong W., Yang C., Jiang W., Tian J, & Gu Y. (2023). Formate as a supplementary substrate facilitates sugar metabolism and solvent production by Clostridium beijerinckii NCIMB 8052. Synthetic and Systems Biotechnology, 8(2), 196-205.

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Profiling miRNA Expression in Ovarian Cancer

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Total RNA was extracted from A2780 and A2780/DDP cells using Trizol (Invitrogen, USA) and miRNeasy mini kit (QIAGEN, Denmark) according to manufacturer’s instructions. The quality and quantity were measured by the spectrophotometer (Beckman Coulter, DU730, USA) and RNA integrity was verified by gel electrophoresis. RNA (1ug) was labeled with Hy3 fluorescent using the miRCURY Power labeling kit (Exiqon, Vedbaek, Denmark) and then hybridized on the miRCURYLNA Array (v.18.0, Exiqon) containing 1887 human miRNA capture probes annotated in miRBase 18.0. Following hybridization, the fluorescent signal images were acquired using a Genepix 4000B scanner (Axon Instruments,Union City, CA, USA) and were analyzed with Genepix Pro 6.0 soft-ware (Axon Instruments), in which the median normalization was obtained. Two-fold or larger change was set as a threshold of significant difference.

Li N., Yang L., Wang H., Yi T., Jia X., Chen C, & Xu P. (2015). MiR-130a and MiR-374a Function as Novel Regulators of Cisplatin Resistance in Human Ovarian Cancer A2780 Cells. PLoS ONE, 10(6), e0128886.

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Quantifying Anthocyanin Content in Seedlings

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Growth light or high light treated seedlings were harvested, weighed, and ground in liquid nitrogen to the fine powder. Anthocyanin measurements were performed as described (Neff and Chory, 1998 (link)). Briefly, the pigments were extracted in methanol with 1% HCl. Water was added and the chlorophyll was extracted with an equal volume of chloroform. Total anthocyanin was determined by measuring the A530 and A657 of the aqueous phase by using a spectrophotometer (DU-730, Beckman). Quantification of anthocyanins was performed using the following equation: Anthocyanin contents = (A530 - 0.25*A657)/weight.

Huang J., Zhao X, & Chory J. (2019). The Arabidopsis Transcriptome Responds Specifically and Dynamically to High Light Stress. Cell reports, 29(12), 4186-4199.e3.

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