Apostle MiniMaxTM High Efficiency cfRNA Isolation Kit

designed for high efficiency cfRNA isolation


Introduction

cfRNAs in biofluids like serum or plasma usually presents as short fragments<1000 nt, and even smaller cell-free miRNA ~20 nt. Although the concentration of cfRNAs is extremely low, cfRNAs are crucial biomarkers for cancer and other diseases. Powered by Apostle MiniMaxTM technology, this cfRNA isolation kit is featured for its efficient recovery of all RNAs in the range between 17-1000 nt, without using phenol or chloroform, and ready for a broad range of subsequent applications, including sequencing, PCR, etc.

Performance data

Apostle MiniMaxTM provides superior RNA isolation efficiency for RNA ladders spiked in biological medium. The isolation efficiency over 90% can be achieved, for both small RNAs (17-150 nt) and RNAs up to 1000nt (Exhibit 1).


MiniMax data image

Exhibit 1. Over 90% RNA recovery in range between 17 – 1000 nt.

RNA (100-1000 nt) and small RNA ladder (17-150 nt) was spiked in serum or plasma, followed by isolation with Apostle MiniMaxTM High Efficiency cfRNA Isolation Kit (red curve), and compared with original RNA ladder (blue curve) using Bioanalyzer 2100 (A) RNA pico kit for 100-1000 nt and (B) small RNA kit for 17-150 nt fragments. Apostle MiniMaxTM High Efficiency cfRNA Isolation Kit offers superior RNA and small RNA recovery efficiency of > 90%.

Apostle MiniMaxTM High Efficiency cfRNA Isolation Kit is suitable for processing plasma collected in various major blood collection tubes (BCTs). Compared to other competitors (a major column-based product), Apostle MiniMax TM High Efficiency cfRNA Isolation Kit provides consistently better performance (1.1-2000 times higher, Exhibit 2).


MiniMax data image

Exhibit 2. 1.1 to 2000 times higher exogenous RNA recovery rate than a column-based major alternative product.

cfRNA was isolated from plasma using Apostle MiniMax TM High Efficiency cfRNA Isolation Kit (red) or a column-based major competitor (green). A synthetic RNA mimic cel-miR-39 was spiked in during the extraction procedure as the reference to evaluate recovery rate. The qPCR amplification plot and CT value show 1.1, 9, and 2000 times higher RNA recovery rate when compared to column-based major alternative product for plasma collected in K3EDTA tube, cfRNA BCT vendor 1, or cfRNA BCT vendor 2, respectively.

The conclusion is further confirmed by endogenous mRNA and miRNAs isolation and qPCR measurements. cfRNA was isolated from 200 uL plasma, using Apostle MiniMaxTM High Efficiency cfRNA Isolation Kit, and a major column based competitor. Blood were collected in K3EDTA BCTs or major specialized cfRNA BCTs and processed to obtain cell-free plasma. Beta-globin, miR-21, miR-U6, and miR-15a were measured by qPCR using the isolated cfRNA, where Apostle MiniMaxTM High Efficiency cfRNA Isolation Kit provides 1.1-480 times higher isolation efficiency compared to major competitor (Exhibit 3-5).


MiniMax data image

Exhibit 3. 1.1 to 2.4 times higher RNA recovery rate than a column-based major alternative product.

cfRNA was isolated from plasma (collected in K3EDTA BCT) using Apostle MiniMaxTM High Efficiency cfRNA Isolation Kit (red) or a major alternative product (green). RNA and small RNA targets including beta-globin, miR-21, U6, and miR-15 were quantified by qPCR amplification plots and CT values. The cfRNA amount isolated using Apostle MiniMaxTM High Efficiency cfRNA Isolation Kit are 1.1 to 2.4 times higher than the column-based alternative product.

Exhibit 4. 3.5 to 480 times higher RNA recovery rate than a column-based major alternative product.

cfRNA was isolated from plasma (collected in cfRNA BCT vendor 1) using Apostle MiniMaxTM High Efficiency cfRNA Isolation Kit (red) or a major alternative product (green). RNA and small RNA targets including beta-globin, miR-21, U6, and miR-15 were quantified by qPCR amplification plots and CT values. The amount of cfRNA isolated using Apostle MiniMaxTM High Efficiency cfRNA Isolation Kit are 3.5 to 480 times higher than the column-based alternative product.
MiniMax data image
MiniMax data image

Exhibit 5. 1.6 to 6 times higher RNA recovery rate than a column-based major alternative product.

cfRNA was isolated from plasma (collected in cfRNA BCT vendor 2) using Apostle MiniMaxTM High Efficiency cfRNA Isolation Kit (red) or a major alternative product (green). RNA and small RNA targets including beta-globin, miR-21, U6, and miR-15 were quantified by qPCR amplification plots and CT values. The amount of cfRNA isolated using Apostle MiniMaxTM High Efficiency cfRNA Isolation Kit are 1.6 to 6 times higher than the column-based alternative product.
Summary

In summary, Apostle MiniMaxTM High Efficiency cfRNA Isolation Kit offers superior isolation efficiency of cell-free RNAs between 17 nt to 1000 nt, without phenol or chloroform. It is suitable for processing samples collected in various major blood collection tubes, especially specialized cfRNA BCTs which will prevent RNA degradation during storage.