RNA & DNA Modification Detection Services

Q: What type of sample can I use?

ELISA: plasma, serum, urine, extracted RNA/DNA Dot blot: plasma, serum, urine, extracted RNA/DNA, other nucleic samples MeRIP/MeDIP: Extracted RNA/DNA (cells/tissues can also be used, however a sample preparation fee will be charged for RNA/DNA extraction) MSP/MSRE: plasma or serum

Q: Do you sell DNA or RNA methylation detection kits that I can use in my lab?

Yes, RayBiotech stocks methylation detection kits for in-lab use. Our catalog includes ELISA kits and dot blot kits for global m6A, m5C, m1A, m7G, ac4C, and A-to-I editing. We also carry gene specific RNA and DNA enrichment kits for m7G, m6A, m1A, m5C.

Our Comprehensive RNA/DNA Research Solutions

To assist you with your studies, RayBiotech offers a complete line of service platforms to examine both RNA and DNA methylation. Our portfolio includes measurement of global changes in m1A, m6A, m5C and m7G through ELISA and dot blot assays, as well as specific gene or region modification detection by methylated RNA immunoprecipitation (MeRIP-qPCR), methylated DNA immunoprecipitation (MeDIP-qPCR), methylation-specific PCR (MSP) and methylation-sensitive restriction enzymes (MSRE) methods.

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What is RNA methylation?

RNA methylation is an epigenetic modification in which methyl groups are added to particular nucleotides within the RNA strand without altering the gene sequence. This modification thereby exerts regulatory control over their functionality. Protein regulators of RNA methylation include “writers” (methyltransferases), “erasers” (demethylases), and “readers” (binding proteins), enzymes which deposit, remove, and recognize methylated RNA, respectively. Among the prevalent RNA methylation modifications, N6-methyladenosine (m6A), 5-methylcytosine (m5C), N1-methyladenosine (m1A) and N7-methylguanosine (m7G) stand out, with m6A being the most prevalent and thoroughly investigated (Fig. 1).

Figure 1. Molecular structures and modification enzymes, including writers, erasers and readers of 4 kinds of RNA methylation.

Role of RNA Methylation

The development of sequencing technology has clarified the pivotal role of RNA modifications in the epigenetic regulation of post-transcriptional gene expression. m6A has been found to affect oocyte maturation and embryonic development as well as the tumorigenic process, including tumor proliferation, invasion, and metastasis by modulating oncogenes and tumor suppressor genes. m5C-related proteins have been identified as participants in embryonic development, plant growth, and neural stem cell differentiation. m1A is also known to be involved in these developmental processes. m7G dysregulation is linked to neurodevelopmental disorders and neurodegenerative diseases.

RNA methylation signatures, particularly m6A, m5C, m1A and m7G, have been suggested as diagnostic and therapeutic targets for various diseases. Despite advances in the structure and pharmacology of RNA methylation regulation that have improved drug discovery, regulating these proteins by various post-translational modifications has received little attention. Therefore, the field of RNA methylation is only in its infancy.

DNA Methylation Detection

DNA methylation is an epigenetic modification by which methyl groups are added to the DNA molecule, primarily at cytosine residues. This modification can have significant alterations on gene expression. For example, when located within a gene promoter, DNA methylation typically acts to repress gene transcription.

Methylation-specific PCR (MSP) is the most commonly used method for gene/sequence-specific detection of DNA methylation for research and clinical applications. Focusing on this method, we offer services using magnetic beads for nucleic acid extraction from urine exosomes, peripheral blood, cells, serum and plasma. Magnetic beads are also used in our DNA bisulfite conversion service to convert unmethylated cytosine residues into uracil for gene methylation analysis in genomic DNA .

Our Platforms for Global RNA/DNA Quantification

RayBiotech’s extensive expertise in immunoassay development has led to the implementation of three precise and efficient detection methods for global m6A, m5C, m1A and m7G modifications: ELISA, dot blot assays and immunofluorescence staining. By directly detecting each modification from RNA isolated from any species, each of our methods can offer researchers insights into the role of RNA methylation in their experimental system.

ELISA Service Features

Absolute quantification from a standard curve
High sensitivity: LOD as low as 0.16 ng/mL
High specificity: no cross-reactivity among the other three modifications (Fig. 2)
Compatible with DNA, RNA, cell culture medium, plasma, serum, urine or other body fluids
Low sample volume requirement: 100 ng of DNA/RNA or 10 µL of plasma, serum, urine or other body fluids
Service report in Excel formats which includes standard curves for each analyte, raw OD values and averages provided. Sample concentrations for each target, including original and calculated (accounting for dilutions)

Figure 2. ELISA standard curves illustrating lack of cross-reactivity between m1A, m6A, m7G and m5C.

Dot Blot Service Features

Semi-quantitative
High sensitivity: as low as 20ng RNA
High specificity: no cross-reactivity among m6A, m5C, m1A and m7G
Compatible with DNA, RNA, plasma, or serum
Low sample volumes: 100 ng of DNA/RNA or 10 µL of plasma, serum, urine or other body fluids
Service report in Excel and PDF formats which includes raw Dot blot images, semi-quantification, original and calculated m1A, m6A, m7G and m5C concentrations (accounting for dilutions) provided

Figure 3. Representative image of m1A-, m6A-, m7G- and m5C-specific dot blot assay of RNA and DNA samples, methylene blue staining is used as a positive control.

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Our Platforms for Gene-Specific RNA Methylation Quantification

RayBiotech’s m1A-, m6A-, and m7G-RNA enrichment and quantification technologies are based on the gold standard method of MeRIP (Methylated RNA Immunoprecipitation) combined with quantitative real-time PCR to identify the abundance and enrichment location of methylation signatures in specific genes. Our streamlined procedure has been validated for RNA methylation enrichment detection in human, mouse, and rat.

MeRIP-qPCR Service Features

High enrichment: True target modification-enriched regions can be reliably identified, and high-resolution mapping achieved
Low input: as low as 500 ng RNA
Service report in Excel and PDF formats which includes the measured Ct (cycle threshold) values from the qPCR assay and the ratio of RNA methylation enrichment based on input as well as a summary of the service background, controls, and procedure

Figure 4. RNA methylation enrichment and quantification by PCR and qPCR. Left panels are representative agarose gels showing 2 transcripts after PCR amplification. Right panels show percentage enrichment from the qPCR assay. 2% Input RNA sample before IP was used as the positive control and IgG-IP was used as the negative control.

Our Platforms for Gene-Specific DNA Methylation Quantification

RayBiotech offers a series of services using magnetic beads for nucleic acid extraction, DNA bisulfite conversion, MSP and MSRE on specific gene DNA methylation analysis for research and clinical applications.

We also offer services for m5C DNA enrichment and quantification based on the gold standard method of MeDIP (Methylated DNA Immunoprecipitation) combined with quantitative real-time PCR to identify the abundance and enrichment location of methylation signatures in specific genes. Our streamlined procedure has been validated for DNA methylation enrichment detection in human, mouse, and rat.

MSP and MSRE-PCR Service Features

High stability, time and labor saved by automated methods
Low input: as low as 50 µL serum or plasma
Service report in Excel and PDF formats which include the measured Ct (cycle threshold) values from the qPCR assay and the relative DNA methylation level as well as a summary of the service background, controls, and procedure

m5C MeDIP-qPCR Service Features

High enrichment: True target modification-enriched regions can be reliably identified, and high-resolution mapping achieved
Low input: as low as 500 ng DNA
Service report in Excel and PDF formats which includes the measured Ct (cycle threshold) values from the qPCR assay and the ratio of DNA methylation enrichment based on input as well as a summary of the service background, controls, and procedure

Figure 5. DNA methylation enrichment and quantification by PCR and qPCR. Left panels are representative agarose gels showing DRD5 and GAPDH transcripts after PCR amplification. Right panel shows percentage enrichment from the qPCR assay. 2% Input DNA sample before IP was used as the positive control and IgG-IP was used as the negative control.

See What Our Gene-Specific RNA/DNA Detection Services Can Do For Your Research

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Frequently Asked Questions

Still have questions?

What type of sample can I use?

How do I ship my samples?

Start by preparing your sample(s) - Tips on Sample Preparation. Then, place each sample into a labeled vial. Freeze the sample right after collection, if possible. You can use a quick freeze procedure. Include a printed copy of the sample service form with your shipment and indicate the quote number or what type of service you’ve requested (ELISA, Dot Blot, MeRIP-qPCR, MeDIP-qPCR, MSP, or MSRE-PCR) on the shipping label. Samples must be free of infections, such as HIV, Hepatitis B, or other communicable viruses, etc.

View more detailed instructions here.

Do you sell DNA or RNA methylation detection kits that I can use in my lab?

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What is RNA methylation?

Figure 1. Molecular structures and modification enzymes, including writers, erasers and readers of 4 kinds of RNA methylation.

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RNA & DNA Modification Detection Services