Table 2 The collections of the current popular IVT-mRNA detection methods.

From: RT-IVT method allows multiplex real-time quantification of in vitro transcriptional mRNA production

Detection methods

Duration

Price/reactiona

Detection components

Instrument

Origins

RT-IVT

3 h (real-time)

3.804 or 0.104b

BFQ probes (Sangon Biotech) PBCV-1 DNA ligase (NEB)

RT-PCR thermocycler or microplate reader

This study

Spectrophotometry

5 h

4.458

RNA purification kit (TaKaRa)

Spectrophotometer

(11)

RT-PCR

8 h

14.117

RNA purification kit (TaKaRa) cDNA Synthesis kit (TaKaRa) SYBR Premix PCR kit (TaKaRa)

RT-PCR thermocycler

(10)

Radioactively tagged nucleotides replacement

12 h

14.860

[α-32P] UTP mixed with ATP, CTP, and GTP (PerkinElmer) Denaturing gel electrophoresis

Radiological safety laboratory and scintillation counter

(12)

  1. aThe cost only applies to the detection components in the reaction and the price was transferred by rate calculation of RMB to USD.
  2. bThe price of 3.804 was calculated using BFQ probes added to PBCV-1 DNA ligase purchased from New England Biolabs (NEB) corporation, while the price of 0.104 was calculated using BFQ probes added to PBCV-1 DNA ligase purified in our laboratory.
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