DNA extraction from peripheral blood samples from the dogs was performed using the DNAeasy commercial kit (Qiagen, Valencia, California, 91355, USA), following the manufacturer's recommendations. The DNA was quantified using a spectrophotometer at 260/280 nm (NanoDrop Technologies ND 1000 UV/VIS, USA), to evaluate the degree of purity, and was then stored at -20 °C.
Nd 1000 uv vis
Manufactured by Thermo Fisher Scientific
Sourced in United States
The ND/1000 UV/Vis is a spectrophotometer designed for measuring the absorbance of samples in the ultraviolet and visible light spectrum. It provides accurate and reliable measurements of sample concentrations and can be used for a variety of applications in the laboratory setting.
Automatically generated - may contain errors
5 protocols using nd 1000 uv vis
1
DNA Extraction from Canine Blood
2
Canine Spleen DNA Extraction
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
DNA extraction from spleen samples from dogs was performed using the commercial DNAeasy® kit (Qiagen, Valencia, California, 91355, United States) according to the manufacturer’s recommendations. The extracted DNA was quantified using a 260/280 nm spectrophotometer (NanoDrop Technologies ND 1000 UV/VIS, United States). The degree of purity was evaluated, and samples were stored at –20°C.
3
Prenatal Triclocarban Exposure and Prefrontal Cortex DNA Methylation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To determine the methylation status of the DNA in the prefrontal cortices of one-month-old mice that had been prenatally exposed to triclocarban, an Imprint Methylated DNA Quantification Kit was used. The methylated DNA was identified by capture and detection antibodies in the following way. First, purified DNA was added to a 96-well plate, allowed to bind, and incubated with the capture antibody. Developing solution was then added, and the wells were monitored for color changes and quantified colorimetrically as previously described [9 (link)]. The amount of methylated DNA was determined spectrophotometrically (ND/1000 UV/Vis; Thermo Fisher NanoDrop, Waltham, MA, USA), and the relative global methylation levels were calculated based on the absorbance of the samples at 450 nm per 50 ng of DNA sample.
4
Transcriptional Response of Cell Lines to Specific Peptides
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After the 24-h exposure to 10 nM of VGVAPG and 10 nM of VVGPGA peptides and in co-treatment with the c-SRC inhibitor I, samples of total RNA were extracted from the analyzed cell lines according to the manufacturer's protocol. The RNA quality and quantity were determined spectrophotometrically at 260 nm and 280 nm (ND/1000 UV/ Vis; Thermo Fisher NanoDrop, USA). Two-step real-time RT-PCR was conducted using the CFX Real Time System (BioRad, USA). The reverse transcription (RT) reaction was performed at a final volume of 20 µL with 800 ng of RNA (as a cDNA template) using the cDNA reverse transcription kit according to the manufacturer's protocol. Products from the RT reaction were amplified using the FastStart Universal Probe Master kit with TaqMan probes as primers for specific genes encoding cMYC, KI67, PPARγ, GAPDH, NFKB2, ELANE, and MTOR according to the manufacturer's protocol. Amplification was carried out in a total volume of 20 µL containing 1.0 µL of the RT product, and GAPDH was used as a reference gene.
5
Quantitative Real-Time PCR of Nuclear Receptor Genes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA was extracted from neocortical cells that were cultured for 7 DIV (approx. 1.5 × 106 cells per sample) using the RNeasy Mini Kit (Qiagen, Valencia, CA) according to the manufacturer’s instructions. The quantity of RNA was spectrophotometrically determined at 260 and 260/280 nm (ND/1000 UV/Vis; Thermo Fisher NanoDrop, USA). Two-step real-time quantitative polymerase chain reaction (qPCR) was performed as previously described [22 (link)]. Both the reverse transcription reaction and qPCR were run on a CFX96 Real-Time System (BioRad, USA). The products of the reverse transcription reaction were amplified using TaqMan Gene Expression Master Mix containing TaqMan primer probes specific to the genes encoding Hprt, Rxrα, Rxrβ, and Rxrγ. Amplification was performed in a total volume of 20 μl containing 10 μl of TaqMan Gene Expression Master Mix and 1.0 μl of reverse transcription product as the PCR template. A standard qPCR procedure was utilized: 2 min at 50 °C and 10 min at 95 °C followed by 40 cycles of 15 s at 95 °C and 1 min at 60 °C. The threshold value (Ct) for each sample was set during the exponential phase, and the delta Ct method was used for data analysis. Hprt was used as a reference gene.
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!