MicroRNA Detection Methods for Mammalian Cell Lines and Their Applications in Therapeutic Protein Production - MicroRNAs as Tools in Biopharmaceutical Production - page 107

Table 7.2 Summary of Three Major Methods for miRNA Detection
Method Sensitivity of detection Normalization Technology maturity Major advantages Major technical challenges

reported controls and applications in

CHO

qRT-PCR Requires the least amount of

total RNA; highest
sensitivity (can detect as low
as 10 copies of synthetic
miRNA)

Small RNA genes
validated

“Gold standard” Most validated technique;

relatively easy to perform;
least demanding in
instrumentation

Low throughput

Microarrays Requires most amount of total

RNA due to labeling
methods, replicates and
controls required
(often > 100

Various in-array
control probes;
dye-swap
technical
replicates

Five years of reported
application and
validation in CHO

High throughput Labor intensive labeling and

hybridization steps; Low
signal to noise ratio

g); lowest
sensitivity and can miss low
abundant miRNA species

μ

NGS Requires somewhat large

amount of total RNA
(1-10

Not reported Very recent ( < 2 years

of application and
validation)

High throughput; sequence
and quantitative data
available simultaneously

Technique under validation
and development

μ g)