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.
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.
A PCR reaction consists of the template DNA, two primers covering the amplification site, an enzyme, and buffers. 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. 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. 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.
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 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.
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