DNA methylation profiling Gene specific profiling SKOV3

A PCR reaction consists of the template DNA, two primers covering the amplification site, an enzyme, and buffers. A quantitative, real-time PCR reaction typically includes all of that plus a probe that can be detected fluorescently as the reaction runs, with no gel required. for detection. However, non-specific product amplification and primer-dimer formation during set-up are major causes of PCR failure. Nevertheless, high-quality DNA polymerase and optimize reaction buffers will certainly lead to a successful PCR reaction.

A PCR reaction consists of the template DNA, two primers covering the amplification site, an enzyme, and buffers. A quantitative, real-time PCR reaction typically includes all of that plus a probe that can be detected fluorescently as the reaction runs, with no gel required. for detection. However, non-specific product amplification and primer-dimer formation during set-up are major causes of PCR failure. Nevertheless, high-quality DNA polymerase and optimize reaction buffers will certainly lead to a successful PCR reaction.

Reporter gene assays are designed to test the regulation of the expression of a gene of interest. This is usually done by linking the promoter of the gene of interest with a gene such as a firefly luciferase, which can be easily detected by addition of luciferin that leads to an enzymatic reaction to produce luminescence. The enzymatic reaction can be correlated to the expression of the gene of interest. Another luciferase gene that can be used is Renilla luciferase. For an appropriate luciferase assay: 1. the reporter should express uniformly in all cells, 2. specifically respond to effectors that the assay intends to monitor, 3. have low intrinsic stability to quickly reflect transcriptional dynamics. It is important to have an equal number of cells plated in each testing condition to avoid any incorrect readouts. Reporter assays could be single or dual reporter assays. The reporter could be both luciferases. Most dual-luciferase assays involve adding two reagents to each sample and measuring luminescence following each addition. Adding the first reagent activates the first luciferase reporter reaction; adding the second reagent extinguishes first luciferase reporter activity and initiates the second luciferase reaction. Dual-luciferase assays have some advantages, including 1. reduces variability, 2. reduces background, 3. normalizes differences in transfection efficiencies between samples.

Isolating DNA from tissues and paraffin-embedded tissue samples can be challenging as double-stranded DNA is physically fragile and highly susceptible to exo- and endonucleases. The best solution is to slice the tissues into smaller pieces and make a homogenate solution (using tissue homogenizer or grinding liquid nitrogen frozen samples) in the presence of DNAse inhibitors. Further, extracting DNA from the nucleus need specific methods by combining physical, mechanical and chemical lysis approaches,

Gene silencing through the use of small interfering RNA (siRNA) has become a primary tool for identifying disease-causing genes. There are several aspects for preparing and delivering effective siRNA to knockdown a target gene. The length of siRNA should be 21–23nt long with G/C content 30–50%. If a validated siRNA sequence for your target gene is not available, use siRNA generated against the entire target gene ORF. Always work with two or three different siRNA constructs to get reliable results. If you are not sure how much siRNA to use for a given experiment, start with a transfection concentration of 10-50 nM and use siRNA-specific transfection reagent to ensure efficient siRNA delivery in a wide range of cells.

Get tips on using DMEM/F12 - Dulbecco's Modified Eagle Medium: Nutrient Mixture F-12 to perform Stem cell culture media Ovarian cancer stem cells (Caov3, 3AO, SKOV3)

DNA ladder is typically used as a reference to estimate the size of unknown DNA samples that are separated based on their mobility in an electrical field. The critical points for running a DNA ladder are compatibility with running buffer, agarose gel percentage, and choosing the correct range of DNA ladder for sizing DNA molecules.

DNA ladder is typically used as a reference to estimate the size of unknown DNA samples that are separated based on their mobility in an electrical field. The critical points for running a DNA ladder are compatibility with running buffer, agarose gel percentage, and choosing the correct range of DNA ladder for sizing DNA molecules.

DNA ladder is typically used as a reference to estimate the size of unknown DNA samples that are separated based on their mobility in an electrical field. The critical points for running a DNA ladder are compatibility with running buffer, agarose gel percentage, and choosing the correct range of DNA ladder for sizing DNA molecules.

DNA damage assay is a standard method for determining in-vivo/in-vitro genotoxicity by measuring the breaks in the DNA chain of animal and plant cells. Initial DNA damage leads to cell cycle arrest and, at the final stages, leads to induction of senescence or cell death (apoptosis, necrosis, autophagy, or mitotic catastrophe). Detection of DNA damage from mild to moderate to severe is challenging when studying genotoxicity in the pool of cells. It is favorable to use DNA damage assay kits available for prominent identification of the extent of damage in the analysis.