Total genomic DNA was extracted from peripheral blood and urine samples using a QIAamp DNA Blood Mini Kit or QIAamp DNA Mini Kit (Qiagen GmbH, Hilden, Germany) according to the manufacturer’s instructions. The concentration and purity of DNA were assessed using a 2000c UV-VIS Spectrophotometer (Thermo Scientific, Wilmington, DE, USA).
2000c uv vis spectrophotometer
Manufactured by Thermo Fisher Scientific
Sourced in United States
The 2000c UV-Vis spectrophotometer is a laboratory instrument that measures the absorption of ultraviolet and visible light by a sample. It is used to quantify the concentration of analytes in a solution by analyzing the amount of light absorbed at specific wavelengths.
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4 protocols using 2000c uv vis spectrophotometer
1
Genomic DNA extraction from blood and urine
2
Microbial Community Profiling in Anaerobic Baffled Reactor
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After 90 d of stable operation, test samples were collected from the biological packing materials in the anaerobic (A-1, A-2, A-3), anoxic (B1, B-2) and aerobic reaction zones (C-1, C-2) of the ABR. The biofilms on the packing materials were washed and collected under sterile conditions. A MO BIO PowerSoil™ DNA isolation kit (Qiagen, Hilden, Germany) was used for DNA extraction according to the method of Wan et al. [28 (link)]. The concentration and purity of the extracted DNA was determined using a 2000c UV-Vis spectrophotometer (Thermo Scientific, USA). Samples (5μL) were separated by 1% agarose gel electrophoresis to isolate the genomic DNA. Pure samples were then amplified by the polymerase chain reaction (PCR) method on an ABI GeneAmp 9700 (Thermo Scientific, USA). Conservative primers 338F (5′-ACTCCTACGGAGGCAGA -3′) and 806R (5′- GGACTACHVGGGGTWTCTA -AT-3′) were used for PCR amplification. The PCR amplification process was performed using the TransStart® FastPfu DNA Polymerase kit based on the procedure of Huang et al. [29 (link)]. The amplified products were purified and sequenced by Shanghai Majorbio Bio-Pharm Technology Co., Ltd. (Shanghai, China). The 16S rRNA gene sequences were compared with the NCBI BLAST® database, and then analyzed according to the sequence analysis method of Hao et al. [30 (link)].
3
Curcumin Encapsulation and Entrapment Efficiency
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The EE% of AuNP@Ng/Cur was determined by measuring the amount of curcumin indirectly from the supernatant. Briefly, Curcumin (0.25 and 0.5 mg/ml) was dissolved in ethanol and then added to both 1 ml Ng and 1 ml AuNP@Ng and stirred for 24 h at room temperature by protecting from light. The following day, nanogel was centrifuged at 3,000 rpm for 5 min. The supernatant was collected, and the amount of excessive curcumin was estimated by using Thermo scientific 2000c UV-Vis spectrophotometer at 450 nm. The EE% and gDL% were calculated using the following equations (Equations 3, 4) (Sarika et al., 2016 (link); Luckanagul et al., 2018 (link)).
4
Cisplatin Modulates Gene Expression in Gastric Cancer Cells
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RT-PCR analysis. The MKN45 and SGC-7901 cells were isolated in the exponential growth phase and seeded in a 96-well plate (5x10 3 cells/well). Cisplatin (1, 2 or 4 µmol/l) or an equal volume of PBS was added to the cells, which were then cultured for 24 h. The cells were then collected and mRNA extraction, cDNA synthesis and PCR amplification were performed according to the manufacturer's instructions of the TRIzol ® and RT-PCR kits. The purity and quantity of RNA were analyzed using a 2000c UV-Vis Spectrophotometer (Thermo Fisher Scientific, Inc.). The following primers were used in the RT-PCR in the present study: p53 forward, 5'-GGTCTCCTCCACCGCTTCTTG TC-3' and reverse, 5'-GGCCTCATCTTGGGCCTGTGT-3'; p53β forward, 5'-GTCACTGCCATGGAGGAGCCGCA-3' and reverse, 5'-ATGGAGGAGCCGCAGTCAGAT-3'; Bax forward, 5'-ACCAAGAAGCTGAGCGAGTGTC-3' and reverse, 5'-ACAAAGATGGTCACGGTCTGCC-3'; β-actin forward, 5'-GAGCCACATCGCTCAGACAC-3' and reverse, 5'-TCGAGGAAACAAATTAAGAA-3'. The lengths of the amplified products were 690 bp for p53, 1050 bp for p53β, 365 bp for Bax, and 539 bp for β-actin. PCR conditions were set as 35 cycles at 94˚C for 5 min, 94˚C for 30 sec, 55-58˚C for 30 sec and 72˚C for 30 sec, followed by 72˚C for 10 min. The PCR products were subsequently analyzed by 2% agarose gel electrophoresis. The results were scanned and recorded by the Biospectrum AC Gel Imaging system.
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