rna-isolation-purification-tissue-mouse-hippocampus

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

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

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

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

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

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.

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

Get tips on using Human C-Reactive Protein/CRP DuoSet ELISA to perform ELISA Human - C-Reactive Protein/CRP

Get tips on using Human CRP/C Reactive Protein PicoKine™ ELISA Kit to perform ELISA Human - C-Reactive Protein/CRP

A PCR reaction consists of the template DNA, two primers covering the amplification site, an enzyme, and buffers. Multiplexing such a reaction amplifies the design challenges where one target requires 3 primers, which should be exclusively bound nowhere in the template DNA or to each other. Similarly, two targets require 6, three require 9, and so on. Each amplicon needs to be either a different size (for gels) or labeled with a different fluorescent tag that is spectrally distinct from the others in the reaction. Further complicating this, different targets in the reaction can compete with each other for resources and causes more challenges in the detection of amplicons. However, with proper primer designing, their validation, optimize quality and concentration of the enzyme and buffers certainly lead to a successful multiplex PCR reaction.