Thiol (SH)-linked alkylation for the metabolic sequencing of RNA, short for SLAM-Seq, is an orthogonal-chemistry-based transcript-mapping method that detects 4-thiouridine (s4U) (a type of nucleoside analog) which incorporates into total RNA at single-nucleotide resolution. SLAM-Seq can be used to map the dynamics of RNA-polymerase-II-dependent gene expression and is an accessible, rapid, scalable, and cost-effective method for studying mechanisms of gene regulation.
SLAM-Seq is a next-generation sequencing (NGS)-based technology for direct quantification of 4-thiouridine (s4U)–labeled mRNAs within the total mRNA pool. This procedure is achieved through the introduction of s4U to thymine and alkylation of the thiol group, which makes modified T deem to be cytosine and induces misincorporation of guanine during reverse transcription instead of adenine, enabling the detection of s4U as thymine-to-cytosine conversion in 3' -end mRNA-sequencing. By sequencing these G mismatches, the newly synthesized mRNA, or other long RNAs and small RNAs can be directly quantified. Integrating rapid chemical-genetic perturbation and SLAM-seq establishes a simple yet powerful strategy for probing specific and global direct functions of transcription factors and cofactors. Meanwhile, SLAM-seq also allows for the study of intracellular RNA dynamics, from transcription, RNA processing to RNA stability when combined with metabolic labeling protocols.
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|High resolution and sensitivity||Long RNA probe length provides high binding affinity to the target lncRNA||Our integrated bioinformatics pipeline can be tailored to suit your project.||Provides one-stop service for library construction, sequencing, sample QC and data analysis.|
quality assessment and quantification.
cDNA library preparation.
Visualize and preprocess results, and perform custom bioinformatics analysis.
RNA sample (concentration ≥ 1 ng/µL, quantity ≥ 5 µg)
1.8 ≤ OD260/280 ≤ 2.2, OD260/230 ≥ 2.0, RIN ≥ 6.5, 28S:18S ≥ 1.0.
Please make sure that the RNA is not degraded nor contaminated.
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, full statistical analysis & alignments, custom analysis reports on customer request.