mRNA Quality Control
mRNA Quality Control
Benchtop PAGE mRNA QC is the New LC/MS
Tired of wait times and costs associated with outsourcing your mRNA for liquid chromatography/mass spectrometry (LC/MS) 5’ capping efficiency and 3’ tail length analyses? Now you can easily perform precise quantitative mRNA 5’ capping efficiency and 3’ poly(A) tail length assessments using standard lab equipment with the EZ-QC™ mRNA Assay Kits.
Discover quantitative fluorescence-based benchtop polyacrylamide gel electrophoresis (PAGE) assays for accurate and cost-effective QC of mRNA 5’ capping and 3’ poly(A) tail length analysis in a single day. Accelerate your mRNA research with the EZ-QC™ Kits for a fraction of the cost of LC/MS.
Benefits:
- Fast benchtop 5’ capping and 3’ tailing QC analyses
- Low-input mRNA (picomoles, not micromoles!)
- Uses common polyacrylamide gel electrophoresis (PAGE) equipment
- Cost-effective – stretch your budget for more experiments
How Our EZ-QC™ mRNA Assays Work
EZ-QC™ mRNA Capping Efficiency Assay Kits
Quantitate total capping efficiency (% capped vs uncapped RNA) of synthesized mRNA
How they work:
- NEW! Now with “EZ-QC™ mRNA Capping Efficiency Targeting Oligonucleotide Designer” tool.
- A chimeric RNA-DNA-RNA Targeting Oligonucleotide hybridizes to the 5′ end of synthesized mRNA.
- The EZ-QC™ XBG mRNA Capping Efficiency Assay Kit supplies an XBG 5′ UTR Targeting Oligo for mRNA with an XBG 5′ UTR.
- The EZ-QC™ mRNA Capping Efficiency Assay Kit requires a user-provided Targeting Oligo for mRNA with any known 5′ UTR sequence.
- The hybridization complex undergoes RNase H cleavage, releasing capped and uncapped 5′-end fragments.
- Released capped and uncapped 5′-end fragments are resolved using PAGE and quantitated by fluorescence-based gel band analysis.
EZ-QC™ mRNA Cap 1 Efficiency Assay Kit
Quantitates the percentage of Cap 1 vs Cap 0 caps of synthesized mRNA
How they work:
- 5′ ends of a synthesized mRNA sample are labeled fluorescently with cyanine5 hydrazide (Cy5).
- Hydrolysis of the Cy5-labeled mRNA mixture releases Cy5-labeled Cap 1 and Cap 0 caps.
- Size resolution by PAGE and fluorescence-based gel imaging is used to determine percentage of Cap 1 vs Cap 0.
- Note: The EZ-QC™ mRNA Cap 1 Efficiency Assay labeling chemistry is not compatible with cap structures that contain a 3′-O-methyl group on the capping G nucleotide such as Anti Reverse Cap Analog (ARCA) or some versions of GpppAG cap analogs.
EZ-QC™ mRNA Poly(A) Tail Length Assay Kit
Quantitates poly(A) tail length of synthesized mRNA
How they work:
- RNase A is used to digest the synthesized mRNA, leaving only intact Poly(A) tails.
- Poly(A) tails are stained with fluorescent dyes (e.g., SYBR Gold Nucleic Acid Gel Stain (Invitrogen) + provided poly(A) fluorescence enhancer).
- Poly(A) tail lengths (from 20 to >300 A’s) are resolved using PAGE with our proprietary Poly(A) 20-mer Ladder and quantitated by fluorescence-based gel band analysis.
