m6A-eCLIP Service: Enhanced meRIP-seq for identifying m6A RNA modification sites

Description

m6A-eCLIP: Enhanced meRIP-seq for superior m6A-seq

Send us your purified RNA for efficient and precise mapping of m6A modification sites

Eclipse BioInnovation´s m6A-eCLIP library preparation for m6A-seq combines the use of an m6A antibody with the eCLIP technology, based on Van Nostrand, Yeo et al. (Nature Methods, 2016 June; 13(6): 508–514.) which results in an enhanced meRIP-seq protocol.

• More sites with less input: The m6A-eCLIP high throughput m6A-seq method enables profiling m6A marks with 100-fold less mRNA input than conventional approaches such as meRIP-seq

• Precise m6A site resolution: Single-nucleotide modification site calling

• Unbiased with high specificity: m6A eCLIP captures the DRACH motif as well as enriched modifications near stop codons and in the 3´UTRs

N6-methyladenosine (m6A) is the most prevalent epitranscriptomic RNA modification found in eukaryotes, and m6A has been shown to regulate many stages of RNA biology including splicing, secondary/tertiary structure, localization, stability, and translation. Adenosine modifications are actively added, recognized, and removed by a series of “writer”, “reader”, and “eraser” proteins, making m6A a dynamic and reversible system for regulating RNAs.

m6A-eCLIP Kit is a targeted RNA modification sequencing method that has applications in many fields of research. Send us your purified RNA to globally map m6A RNA modification states, which are associated with cancer, autoimmune disease, obesity, and other disease indications, e.g. acquisition of host-like m6A by viral RNA also plays a role in evading detection by the innate immune system.

m6A-eCLIP Service Description: 

You provide: 50ug Total RNA; RIN >7, Isolated from Cells or Tissues

The service comprises:

• Poly(A) selected and fragmentation of provided RNA.
• Binding of the Eclipse m6A antibody to m6A sites.
• UV crosslinking of m6A antibody to mRNA.
• Immunoprecipitation
• Library preparation
• Sequencing: 50M reads per sample and subsequent comprehensive data analysis

Deliverable:

You receive the following comprehensive Data Analysis Package:

⋅ RAW SEQUENCING DATA | FASTQ:

Reads directly off sequencer and demultiplexed per sample for data back-up and publication

⋅GENOME ALIGNMENTS| BAM:

Reads filtered of repetitive elements, aligned, and PCR deduplicated for additional downstream analysis

⋅COVERAGE TRACKS| BIGWIG:

Normalized read coverage on positive and negative strands for visualization in a genome viewer such as IGV

⋅SCORED PEAKS & SITES| BED:

Genomic regions (aka “peaks”) and single nucleotide positions scored after input normalization as putative m6A sites for target exploration. Includes log2 enrichment scores and p-values 

-SUMMARY REPORT | HTML:

Interactive report providing an overview of m6A-eCLIP results

For an example report click here

We are offering scaled pricing depending on the number of RNA samples provided –
please contact us for a quote!