Photo-crosslinking-assisted m6A sequencing (PA-m6A-seq) is a next-generation sequencing (NGS)-based method to comprehensively detect m6A. RNA methylation modification exists in various transcripts. It can map m6A in sample transcriptome with high precision. Our PA-m6A-seq service is dedicated to detecting and analyzing m6A RNA modification in a sample, providing a powerful tool for research of intracellular translation, regulation, and protein modifications.
N6-methyladenosine (m6A), methylation modification on the 6th nitrogen atom of RNA adenine, is one of the most abundant chemical modifications on eukaryotic mRNA. As a dynamic and reversible modification method, m6A is mainly enriched in the promoter region and termination codon region of mRNA. It plays an important role in regulating gene expression, splicing, RNA editing, RNA stability, controlling mRNA longevity and degradation, and mediating circular RNA translation is a hot spot in the epigenetic transcriptome, in which accurate identification of the m6A site will provide insights into its biological functions and mechanisms. We provide integrated PA-m6A sequencing service, which can map m6A in sample transcriptome with high precision. The technique synthesizes 4-thiouracil (4SU) into RNA by adding 4SU into the growth medium. After m6A immunoprecipitation, recovered m6A-containing RNA is cross-linked to the anti-m6A antibody under 365-nm ultraviolet light. Later, RNase T1 digests the cross-linked RNA to approximately 30nt and performing efficient sequencing. PA-m6A-seq at single-base resolution can determine the single consensus methylation sequences, including known and new m6A sites. In the meantime, this ultraviolet cross-linking strategy effectively provides insight into m6A-containing RNA and RNA-binding proteins.
|Single-base Resolution||High Efficiency||Professional Team||Bioinformatics Service|
|Single-base resolution allows accurate detection of accurate sites of m6A.||Adopts carefully optimized experimental procedure, achieving high efficiency and specificity.||Provides various in-depth data analysis to satisfy customer needs.||Provide standard and customized bioinformatics analyses, which are reflected in the report.|
quality assessment and quantification.
RNA fragmentation; immunoprecipitation;
cross-linked reaction; m6A library preparation.
Visualize and preprocess results, and perform custom bioinformatics analysis.
RNA sample quantity ≥ 300 ug.
Please make sure that the RNA is not significantly degraded.
Sample storage: RNA can be dissolved in ethanol or RNA-free ultra-pure water and stored at -80°C. RNA should avoid repeated freezing and thawing.
Shipping Method: When shipping RNA samples, the RNA sample is stored in a 1.5 mL Eppendorf tube, sealed with sealing film. Shipments are generally recommended to contain 5-10 pounds of dry ice per 24 hours.
Deliverable: FastQ, BAM, coverage summary, QC report, custom bioinformatics analysis.