RNA extraction from subcutaneous adiposetissue (scAT) in human obesity - Bertin Instruments

Other

RNA extraction from subcutaneous adiposetissue (scAT) in human obesity

Sources: INSERM UMRS U872 (Eq7) Nutriomic, Nutrition andEndocrinology Department, Pitié-Salpêtrière Hospital, Paris, France. (V. Pelloux)

Context

Adipose tissue is a critical player in obesity-related metabolic functions. Indeed, adipose tissue is no longer considered to be simply a passive lipid reservoir, but rather an endocrine organ capable of secreting factors that profoundly influence processes such as feeding behavior, energy flux, and immuno-inflammation. As such, obtaining adipose tissue samples are paramount to the understanding of human obesity. Much of our recent understanding regarding the role of subcutaneous adipose tissue (scAT) in human obesity is a consequence of using modern molecular biology tools, such as the microarray. [1] These techniques are highly dependent on the quality of total RNA obtained. Biopsies of scAT are precious and often small amounts. To improve performance and get a better quality of extracted RNA, we compared two mashing methods.

 

[1] Mutch, D. M. & Clement, K. Unraveling the genetics of human obesity. PLoS Genet 2, e188 (2006).

Results

Yield for both extractions was determined and shown in figure1. The heterogeneity of the amount of RNA obtained was unequal to the weight of the biopsies. An increase in the amount of extracted RNA is observed with Precellys®24. Same extracted RNA purity is observed with these two methods (Figure 2a, 2b). By quantitative PCR, we observed an amplification factor of the leptin gene using Precellys®24, which will quantify the expression of genes weakly expressed in this tissue.

 

extracted RNA yield obtained with Ultraturax and Precellys methods.

extracted RNA yield obtained with Ultraturax and Precellys methods.

 

a) sample 7 manual –RIN 7.0; b) sample7 Precellys RIN 7.

a) sample 7 manual –RIN 7.0; b) sample7 Precellys RIN 7.

ANY QUESTIONS? CONTACT AN EXPERT

Please fill in this form if you wish to contact our sales team or receive further information.

My Account
Forgot password ?

Add a question !

Add a question