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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.

DNA PCR Conventional / Qualitative PCR mammalian DNA

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.

DNA PCR Conventional / Qualitative PCR bacterial DNA

Get tips on using FastDigest RsaI to perform Restriction Enzymes RsaI / AfaI

Products Thermo Fisher Scientific FastDigest RsaI
RsaI R6371 Product

Get tips on using RsaI R6371 to perform Restriction Enzymes RsaI / AfaI

Products Promega RsaI R6371

Get tips on using RsaI NEB#R0167 to perform Restriction Enzymes RsaI / AfaI

Products New England BioLabs RsaI NEB#R0167

Get tips on using AfaI (RsaI) restriction enzyme to perform Restriction Enzymes RsaI / AfaI

Products Takara Bio Inc AfaI (RsaI) restriction enzyme

Get tips on using RsaI (10 U/µL) to perform Restriction Enzymes RsaI / AfaI

Products Thermo Fisher Scientific RsaI (10 U/µL)

Get tips on using RCAS1 siRNA (h) to perform siRNA / miRNA gene silencing Human - ES2 RCAS1

Products Santa Cruz Biotechnology RCAS1 siRNA (h)

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.

Cellular assays Reporter gene assay β-galactosidase substrates SK-Hep-1

Get tips on using pUC8CVX-RsaAΔ0–222 to perform Protein Expression Prokaryotic cells - C. crescentus JS1014 RsaAΔ0–222

Products John Smit, Department of Microbiology and Immunology, University pUC8CVX-RsaAΔ0–222

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