White Paper

Gene Selection Using PrimePCR Plates And CFX Maestro Software

Asain Woman Scientist

Quantitative real-time PCR is one of the most important methods for rapid and reliable quantification of gene expression. In order to perform an accurate analysis of relative gene expression in different samples, tissues, or experimental conditions, reference genes must be used to normalize variation across all samples. While the importance of choosing the appropriate reference genes has been widely recognized, researchers often rely on knowledge over experience and often simply choose to use classical reference genes (GAPDH, B2M, ACTB, TBP, and the like). Although these commonly used reference genes may be stable in many situations, it has been observed that their expression can vary considerably depending on species, genotype, developmental stage, experimental treatment, or circadian rhythm (Jain et al. 2018Lacerda et al. 2014Radonic et al. 2004). Additionally, reference genes reported in the literature for a particular sample type should be validated experimentally prior to use. Some sources of variability that must be included in the reference gene stability survey include sample type and treatment(s). Other experimental sources of variability can be minimized by making sure to harvest cells, prepare and quantitate RNA, and synthesize cDNA using the same methods and reagents. It is important to evaluate several reference genes and select only those that are stably expressed across all samples and sets of conditions. Use of multiple stably expressed reference genes for normalization can further reduce gene expression (Vandesompele et al. 2002).

VIEW THE WHITE PAPER!
Signing up provides unlimited access to:
Signing up provides unlimited access to:
  • Trend and Leadership Articles
  • Case Studies
  • Extensive Product Database
  • Premium Content
HELLO. PLEASE LOG IN. X

Not yet a member of Cell & Gene? Register today.

ACCOUNT SIGN UP X
Please fill in your account details
Login Information
I'm interested in newsletter subscriptions.
ACCOUNT SIGN UP

Subscriptions

Sign up for the newsletter that brings you the industry's latest news, technologies, trends and products.

You might also want to: