What are good housekeeping genes for qPCR?
Most stable genes were Atp5f1, Pgk1 and Gapdh, while the least stable genes were Rps18, Hprt, Tbp and Actb. The results were validated by a proof-of-point qPCR experiment with pluripotent markers Nanog, Rex1 and Oct4 normalized to the best and the worst reference gene identified by the analyses.
What is the purpose of the housekeeping gene in RT PCR?
Housekeeping genes are cellular maintenance genes which regulate basic and ubiquitous cellular functions. In many RT-qPCR reactions, these genes are used as internal control genes without proper validation.
What is considered housekeeping gene?
In molecular biology, housekeeping genes are typically constitutive genes that are required for the maintenance of basic cellular function, and are expressed in all cells of an organism under normal and patho-physiological conditions. The origin of the term “housekeeping gene” remains obscure.
What are the examples of housekeeping genes?
Examples of housekeeping genes include actin, GAPDH and ubiquitin. Compare: luxury gene. See also: genetic expression, transcription.
How do you choose endogenous control?
Choosing and validating an endogenous control
- Select experimental conditions that are representative of your study, e.g. a specific range of cell types, treatments or time points.
- Purify the RNA from all your samples across different test conditions using the same method.
What makes a good housekeeping gene?
The definition of a good housekeeping gene, is one that has stable expression in varying conditions, thus can be used to normalize other influences which may skew the result.
How does housekeeping gene work?
Housekeeping genes are typically constitutive genes that are required for the maintenance of basal cellular functions that are essential for the existence of a cell, regardless of its specific role in the tissue or organism. On the one hand they represent the minimal set of genes required to sustain life.
Is 16S a housekeeping gene?
generally, so-called “house-keeping” genes involved in basic cellular functions are used for normalisation (16S, tus, rpoD, glyA, dnaB, gyrA, pykA/F, pfkA/B, mdoG, arcA).
Is B2M a good housekeeping gene?
In addition, B2M, which has been found to be a stable housekeeping gene in other biological systems [14, 17] showed one of the highest levels of variation in this tissue. A large number of studies have been carried out concerning the validation of housekeeping genes in many different tissues and cell types.
How do housekeeping genes work?
Housekeeping genes are typically constitutive genes that are required for the maintenance of basal cellular functions that are essential for the existence of a cell, regardless of its specific role in the tissue or organism.
What’s endogenous control?
An endogenous control gene is a gene whose expression level should not differ between samples, such as a housekeeping or maintenance gene.
What are endogenous reference genes?
Endogenous reference genes are commonly used to normalize expression levels of other genes with the assumption that the expression of the former is constant in different tissues and in different physiopathological conditions.