A Practical IVT mRNA QC Workflow
From transcription and purification to RNA size, dsRNA impurity and residual T7 RNA polymerase.
Producing an IVT RNA transcript is only the beginning.
Once an mRNA or self-amplifying RNA (saRNA) construct has been transcribed, researchers may need to answer several different quality-control questions.
No single analytical method answers all of these questions. A practical RNA QC workflow therefore combines complementary approaches.
IVT RNA QC at a Glance
T7 RNA Polymerase
RNA Ladder
J2-based ELISA
T7 Polymerase ELISA
Start With the QC Question
| Question | Approach | Information |
|---|---|---|
| Is the RNA the expected size? | RNA Ladder | RNA size |
| Does the RNA appear intact? | Electrophoresis | Band pattern |
| How much dsRNA is present? | dsRNA ELISA | Residual dsRNA |
| Is T7 RNA polymerase still detectable? | T7 RNA Polymerase ELISA | Residual process enzyme |
1. RNA Size & Integrity
RNA electrophoresis provides a practical way to compare an IVT RNA sample with RNA fragments of known sizes.
The appropriate ladder depends on the expected transcript length.
Conventional IVT RNA
Long mRNA
Long RNA / saRNA
Ultra-long RNA
12K High Range RNA Ladder — CR00005-50UL
Nine ssRNA reference bands: 0.2 / 0.5 / 1 / 2 / 3 / 4 / 6 / 9 / 12 kb.
Shop 12K RNA LadderCompare 6K, 9K, 12K and 15K RNA ladders.
RNA Ladder Selection Guide →2. dsRNA Impurity
RNA size alone does not tell researchers how much double-stranded RNA may be present in an IVT preparation.
dsRNA therefore represents a separate analytical question.
Double-Stranded RNA ELISA Kit
J2-based sandwich ELISA for quantitative dsRNA measurement using a standard curve.
C13005-K01Find dsRNA ELISA
Learn why dsRNA is monitored separately from RNA size.
Why dsRNA Impurities Matter →3. Residual T7 RNA Polymerase
T7 RNA polymerase is commonly used to generate RNA during in vitro transcription.
Because the enzyme is part of the production process rather than the intended RNA product, researchers may also choose to evaluate residual T7 RNA polymerase after purification.
T7 RNA Polymerase ELISA Kit
Research-use ELISA for quantitative evaluation of residual T7 RNA polymerase in IVT RNA workflows.
C13004-K01Find T7 RNA Polymerase ELISA
Three Assays. Three Different Questions.
Size
Is the RNA approximately the expected length?
RNA Impurity
How much residual dsRNA is present?
Process Residual
Is T7 RNA polymerase still detectable?
Build Your IVT RNA QC Workflow
Explore tools for long RNA, mRNA and saRNA research — from RNA size assessment to dsRNA and residual process enzyme analysis.
Explore RNA QC Solutions Shop 12K RNA LadderFor Research Use Only. This article provides general information about research-use IVT RNA QC workflows. Refer to the applicable product documentation for specifications and experimental conditions.