Full-upgrade Integrated Service of MeRIP-Seq/ m6A -Seq

Advantages

1. Platform: from MeRIP experiment, library construction, sequencing, analysis to MeRIP-qPCR
2. High Success Rate: successfully delivered more than 1,000 samples and support ng level RNA
3. Flexibility: various library construction methods and fragment sizes
4. Good Repeatability: assured experimental repeatability by rich project experience and comparative analysis among multiple samples

Sample Preparation

1. Tissue: It is suggested to be more than 3g. Partial tissues with less RNA may require more tissues.
2. Cell: It is suggested to be more than 2×107. Partial cells with less RNA may require more cells.
3. Total RNA: It is suggested to be more than 100 μg.

Recommended Sequencing Model and Data Volume

illumina SE50, 20M clean reads
illumina PE150, 6G clean data

Research Approach

illumina SE50, 20M clean reads
illumina PE150, 6G clean data

Peak motif Analysis

motif Detection

For more information, please contact our sales representative.

How to request for the integrated service of MeRIP-seq (m6A)?

Please visit https://www.ribobio.com/en/, take 3 minutes to register an account and place online request for the integrated service of MeRIP-seq.

1. Once the account registration is approved, please clickhttps://www.ribobio.com/en/our-services/to land on the “Request for Services” page.。
2. Choose the service you need, such as MeRIP-seq and then click “High-throughput Sequencing”.
3. Fill in the form and submit. Select the sequencing type MeRIP-seq, complete the sample information, etc. and directly submit successfully.

Research Background of m6A

m6A modification commonly exists in mRNA and various of non-coding RNAs that participates in researches of biological function and mechanism of actions including tumorigenesis and metastasis, embryogenesis, DNA damage repair and virus infection, etc by affecting RNA mature processing, stability, mRNA transportation and translation, RNA editing, etc.
Comparing with the earlier research on DNA and histone epigenetic modification, recently, there exists similar regulation mechanism in RNA. RNA modification has exceeded 150 kinds. N6-methyladenosine (m6A), as one of the most common post-transcriptional modifications, is highly conserved in eukaryote for mediating over 80% of RNA methylation. This methylated modification is very universal, which its functions are mediated by “Writer, Eraser and Reader”.

RNA N6‐methyladenosine methyltransferase‐like 3 promotes liver cancer progression through YTHDF2‐dependent posttranscriptional silencing of SOCS2[J]. Hepatology, 2018, 67(6):2254.

Writer (methyltransferases) catalyzes the development of m6A methylated modification fromRNA in vitro and in vivo, which includes METTL3, METTL 14 and WTAP, VIRMA and RBM15. Erasers (demethylase) are able to remove signal of RNA methylated modification including ALKBH5 and FTO. Readers can identify RNA methylated modification and participate in the process of RNA translation, stability, splicing and transfer, etc, which includes YTHDF1, YTHDF2, YTHDF3,YTHDC1 and YTHDC2.

References

1. Cui Q, Shi H, Ye P, et al. Knockdown of METTL3 or METTL14, key components of the RNA methyltransferase complex, dramatically promotes human GSC growth, self-renewal, and tumorigenesis.
2. Ma J Z, Yang F, Zhou C C, et al. METTL14 suppresses the metastatic potential of hepatocellular carcinoma by modulating N6 -methyladenosine-dependent primary MicroRNA 126 processing.
3. Li Z, Weng H, Su R,et al. FTO, as the first N 6-Methyladenosine RNA Demethylase, can affect the RNA binding of splicing factor SRSF2 and further regulate the splicing and processing procedure of pre-mRNA.
4. Zhao B S, Wang X, Beadell A V, et al. YTHDF2 can identify the modified mRNA and mediate m6A-dependant maternal mRNA clearance.
5. Yang X, Laurent B, Hsu C H, et al. RNA m6A methylation modification, regulated by the methyltransferase METTL3 (methyltransferase-like 3) and the demethylase FTO, serves as a beacon for the rapid recruitment of Pol κ to damage sites to facilitate repair and cell survival.
6. Yang Y, Fan X, Mao M, et al. The consensus m6A motifs are enriched in circRNAs and a single m6A site is sufficient to drive translation initiation. This m6A-driven translation requires initiation factor eIF4G2 and m6A reader YTHDF3.

More to discover……