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Extraction of RNA and microRNA from human milk (623.3)
Author(s) -
Hassiotou Foteini,
Alsaweed Mohammed,
Hepworth Anna,
Lefevre Christophe,
Hartmann Peter,
Geddes Donna
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.623.3
Subject(s) - rna , microrna , rna extraction , small rna , extraction (chemistry) , chemistry , computational biology , chromatography , biochemistry , biology , gene
Human milk contains both free and encapsulated RNA and microRNA in its cell, lipid and skim milk fractions. Recently, the microRNA content of human milk has attracted much attention due to its high variability and the presence of microRNAs with known immunomodulatory functions. However, optimization of the extraction of RNA and microRNA from different fractions of human milk, which is a prerequisite for investigating the functions of these milk molecules, has not been carried out. In this study, we compared eight commercially available kits for their efficacy in extracting RNA and microRNA from the three fractions of human milk. The highest quantities of both RNA and microRNA were recorded for the cell and lipid fractions, and the lowest for skim milk. The most efficient method for RNA and microRNA extraction was the column‐based phenol‐free method, and the least efficient the phenol‐based guanidine method. Interestingly, some kits were more efficient in extracting high quantities of RNA or microRNA, whilst others performed better in extracting high‐purity RNA or microRNA. This now provides guidance for extracting RNA and microRNA from different breastmilk fractions allowing a more comprehensive profiling of these breastmilk molecules and their functions for the breastfed infant.