Option | mRNA QC Test | TAT, days | Amount Required | Method |
---|---|---|---|---|
1 | PolyA Tail Length Assay | 7 | 300pmol | RNase digestion and purification to recover the polyA tail fragment, followed by polyA tail length analysis by small RNA bio-analyzer kit assay or LC-MS. |
2 | Capping Efficiency Assay | 14-21 | 300pmol | RNase digestion to isolate a short oligo from the 5’ end of the mRNA sample to be analyzed by LC-MS to calculate the capping efficiency. |
3 | dsRNA Residue Assay | 10 | 40ug | Slot-blot assay using a dsRNA-specific antibody to test for dsRNA residues in mRNA sample. |
4 | RNA Capillary Electrophoresis | 5 | 10ug | Bioanalyzer assay to test RNA length and purity. |
5 | Total Protein Residue Assay | 7 | 200ug | Nano-orange assay to test the residual protein amount in mRNA sample. |
6 | Plasmid DNA Residue Assay | 14 | 15ug | q-PCR assay to test the residual plasmid DNA residue amount in mRNA sample. |
7 | Quantitative Endotoxin Assay | 5 | 15uL | GenScript ToxinSensor™ Chromogenic LAL Endotoxin Assay to test endotoxin level in mRNA sample. |
8 | BioBurden Assay | 14 | 250uL | TSA plate to recover a broad range of microorganisms such as aerobic and anaerobic bacteria from liquid. |
9 | Sanger Sequencing | 7 | 5ug | RT-PCR and sanger sequencing, alignment with target sequence for data analysis. 2-4X coverage. |
10 | NGS | 14 | 5ug | Illumina platform with 1G data amount, 10,000X depth. |
11 | RNA Aggregation Assay | 7 | 30ug | SEC-HPLC method |
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