Snap-frozen tissue was proteinase K digested, and genomic DNA (gDNA) was extracted using the Blood & Cell Culture DNA Mini kit (Qiagen, Germantown, MD, USA) as indicated. Isolated gDNA was quantified using the BioTek plate reading spectrophotometer (BioTek Instruments, Winooski, VT, USA). gc distribution in diploid cells were detected and quantified by qPCR using Applied Biosystems 7500 Real-Time PCR Systems with TaqMan PCR master mix reagents (Applied Biosystems, Woburn, MA, USA) and transgene-specific primer/probes.
Plate reading spectrophotometer
Manufactured by Agilent Technologies
Sourced in United States
The Plate reading spectrophotometer is a versatile instrument used to measure the absorbance or transmittance of light by samples in microplates. It is designed to quantify the concentration of a substance in a solution by analyzing the interaction of light with the sample.
Automatically generated - may contain errors
Lab products found in correlation
Pierce ldh cytotoxicity assay kit, by Thermo Fisher Scientific (2 mentions)
Taqman pcr master mix reagents, by Thermo Fisher Scientific (1 mentions)
Blood cell culture dna mini kit, by Qiagen (1 mentions)
7500 real time pcr system, by Thermo Fisher Scientific (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Beyotime (1 mentions)
4 protocols using plate reading spectrophotometer
1
Genomic DNA Extraction and Quantification
2
Quantifying Cell Proliferation via MTT Assay
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To measure cell proliferation, an MTT assay was conducted, as described previously.39 Cells (3 × 104 cells/well) were seeded in a 96‐well plate and cultivated for 48 h before the MTT assay. MTT (Beyotime) (20 µL) was added to each well and the cells were cultured for 4 h at 37°C. Finally, a plate reading spectrophotometer (BioTek Instruments, Inc.) was used to measure the absorbance at 570 nm. Experiments were independently repeated three times.
3
Cytotoxicity of Au NCs in Breast Cancer Cells
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MCF-7 and MDA-MB-231 cells were seeded on a 96-wellplate (TPP, Trasadingen, Switzerland) at a density of 1.5·103 cells/well. After 24 h, the old medium was replaced with a fresh medium containing 2.75, 5.5, 13.75 or 27.5 mg/mL Au NCs Rh+/Rh−, while media alone without Au NCs was a control. Cells were incubated for 24 h in the dark. The next day Pierce LDH Cytotoxicity Assay Kit (Pierce Biotechnology, Thermo Scientific, USA) was used to detect extracellular appearance of lactate dehydrogenase (LDH). The concentration of extracellular LDH was quantified by measuring absorbance at 490 and 630 nm wavelength with plate-reading spectrophotometer (BioTek, Winooski, VT, USA). After obtaining absorbance values, they were recalculated as percentage values of cytotoxicity, according to the protocol. For better data representation estimated cytotoxicity (%) was recalculated to viability of cells (100 %—cytotoxicity (%)). Data are expressed as mean ± standard deviation (SD). The statistical significance of differences between studied groups was assessed using a two-tailed independent Student’s t-test at the 95% confidence level. Significance was represented as p-value < 0.05.
4
Cytotoxicity of Tc-99m Labeled BSA-Au NCs
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HEK-293T cells were seeded on a 96-wellplate (TPP, Switzerland) at a density of 1.2 × 103 cells/well. After 24 h, the old medium was replaced with a fresh medium containing 1.5, 3, 7.5 or 15 mg/mL 99mTc-BSA-Au NCs, while medium alone without 99mTc-BSA-Au NCs was a control. Cells were incubated for 24 h in the dark. The next day, Pierce LDH Cytotoxicity Assay Kit (Pierce Biotechnology, Thermo Scientific, Eugene, OR, USA) was used to detect extracellular appearance of lactate dehydrogenase (LDH). The concentration of extracellular LDH was quantified by measuring absorbance at 490 and 630 nm wavelength with plate-reading spectrophotometer (BioTek, Winooski, VT, USA). After obtaining absorbance values, they were recalculated according to the protocol to represent percentage values of cytotoxicity. For better data representation, estimated cytotoxicity (%) was recalculated to viability of cells (100%—cytotoxicity (%)). Data are expressed as mean ± standard deviation (SD). The statistical significance of differences between studied groups was assessed using a two-tailed independent Student’s t-test at the 95% confidence level. The level of statistical significance was expressed as p-values < 0.05.
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