Table 1 Strengths and weaknesses of different approaches used for miRNA measurement
From: The potential of microRNAs in cancer diagnostic and therapeutic strategies: a narrative review
Methodologies | Strengths | Weaknesses |
|---|---|---|
Northern blotting (Hybridization-based approach) | Could identify the lengths of the targeted miRs Canonical method for validating new miRs High specificity | Demand substantial quantities of RNA Low sensitivity Complicated procedure Could identify only one miR at a time Difficult to quantify |
qRT-PCR (Amplification-based detection) | Rapid, high sensitivity and specificity Required a minimal quantity of RNA | The length of the targeted miRs could not be determined Designing primers and probes with precision can be a challenging process Relative quantification Could identify only one miR at a time |
Digital PCR (Amplification-based detection) | Rapid, high sensitivity and specificity Required a minimal quantity of RNA Absolute quantification | The length of the targeted miRs could not be determined Designing primers and probes with precision can be a challenging process Chips and reagents are costly Simultaneously detect and quantify specific miR |
Microarray (Hybridization-based approach) | Simultaneously detect and quantify a panel of miRs | Demand substantial quantities of RNA Only quantify known miRs Low sensitivity and specificity |
Next-generation sequencing (Sequencing-based detection) | Simultaneously detect and quantify a panel of miRs Request an acceptable quantity of RNA High sensitivity and specificity | Complicated procedure The process of library preparation is intricate and can contribute to considerable bias |