Evaluation of five column‐based isolation kits and their ability to extract miRNA from human milk

Abstract

MicroRNA can be found in various body fluids, including breast milk. MicroRNA may be transferred from mother to infant via breast milk and potentially regulate the development of the infant's immune system on a post‐transcriptional level. This study aimed to determine the microRNA extraction efficiency of five RNA extraction kits from human skim milk samples. Their efficiency was determined by comparing microRNA concentrations, total RNA yield and purity. Furthermore, hsa‐miR‐148a‐3p expression and the recovery of an exogenous control, cel‐miR‐39‐3p, were quantified using qPCR. Each kit extracted different amounts of microRNA and total RNA, with one kit tending to isolate the highest amount of both RNA species. Based on these results, the extraction kit ReliaPrep™ miRNA Cell and Tissue Miniprep System from Promega was found to be the most appropriate kit for microRNA extraction from human skim milk. Moreover, further research is needed to establish a standardized protocol for microRNA extraction from breast milk.

Advances in the protection of intestinal mucosal barrier function by milk-derived miRNAs

“…13 Besides, in milk exosomes, researchers have also detected the presence of miRNAs in milk components such as milk fat and whey of bovine milk or human milk, respectively. 14,15…”

“…13 Besides, in milk exosomes, researchers have also detected the presence of miRNAs in milk components such as milk fat and whey of bovine milk or human milk, respectively. 14,15 However, it is so far unclear how mature miRNAs are selectively wrapped/loaded into exosomal vesicles, which may depend on the surface molecules of the exosome membrane, the specific proteins required for transport, and the specific binding motifs of the miRNAs themselves. For example, Shurtleff et al (2016) suggested that the RNA-binding protein Y-box protein I may play an essential role in the secretion of miRNAs into exosomes by HEK293T cells.…”

Evaluation and Standardization of RNA Extractions with Quality for RNA-Seq for Balamuthia mandrillaris

“…In RNA, absorbance at A260 primarily measures nucleic acids, indicating the RNA concentration; absorbance at A280 detects proteins; and absorbance at A230 identifies organic compounds. Also, RNA has a higher 260/280 ratio due to the ratio of Uracil (4.00) compared to that of Thymine (1.47) [17]. This is validated through spectrophotometry using the 260/280 ratio, with an optimal value above 2.0 for RNA [18], and the 260/230 ratio, with an optimal value of 2.2 [19].…”

“…This is validated through spectrophotometry using the 260/280 ratio, with an optimal value above 2.0 for RNA [18], and the 260/230 ratio, with an optimal value of 2.2 [19]. Values above 2.0 for the A260/280 ratio indicate the absence of protein contamination, and values above 2.0-2.2 for the A260/230 ratio indicates the absence of solvent contamination and denote the isolation of high-quality, non-contaminated, or pure RNA [17,20].…”

Role of microRNAs in Immune Regulation with Translational and Clinical Applications

“…On the other hand, recent reports on methodology of miRNA extraction demonstrated higher efficiency of column-based isolation kits compared to Trizol [257,258]. However, different kits apply nonidentical protocols, resulting in variable RNA yield and purity [259,260].…”