BS-seq reads 5-methylcytidine (m⁵C) on tRNA through bisulfite chemistry. Unmodified cytidine is converted and reads as thymine, while m⁵C is protected and stays cytosine, so every modified site is resolved and quantified at a single nucleotide.
Converting cytidine chemistry into an m⁵C map
On tRNA, m⁵C reinforces the folded L-shaped structure and fine-tunes anticodon–codon decoding. Bisulfite conversion deaminates unmodified cytidine to uracil, which is read as thymine, while 5-methylcytidine resists the reaction and is read as cytosine, so the conversion pattern itself carries the position information.
Arraystar Single-Nucleotide tRNA m⁵C Modification Seq(BS-seq) is an end-to-end sample-to-data service for tRNA, from RNA sample QC and tRNA treatment through library construction, sequencing and bioinformatics. Sites are localized within mature tRNA transcripts with relative coordinates, gene attribution, anticodon and isotype, and structural-domain context.
BS-seq is an antibody-free, base-resolution method for 5-methylcytidine on tRNA. Bisulfite conversion turns unmodified C into U, which reads as T, while m⁵C is protected and reads as C, so each m⁵C site is mapped and quantified at single-base resolution.
Standard tRNA m⁵C project — custom designs and add-on analyses available on request
| Service Name | Price |
|---|---|
| Single-Nucleotide tRNA m⁵C Modification Seq(BS-seq) |
What bisulfite sequencing delivers for tRNA m⁵C
Unmodified cytidine becomes thymine while m⁵C stays cytosine, so each modified site is separated from the unmodified background base by base.
The proportion of unconverted reads at a position yields the m⁵C fraction, which supports quantitative comparison between samples.
Bisulfite chemistry avoids enrichment bias and cross-reactivity, which is an advantage inside compact and heavily folded tRNA.
Each call carries relative coordinates, host tRNA gene, anticodon, isotype and structural-domain localization.
Sites can be grouped by structural domain, so acceptor-stem and loop-region m⁵C can be compared directly.
QC, tRNA treatment, library construction, sequencing and bioinformatics are delivered as one project.
m⁵C, tRNA architecture and decoding
In tRNA the modification stabilises the folded L-shaped structure and fine-tunes anticodon–codon decoding, so base-resolution mapping within maturation loops and the anticodon is what distinguishes one site from another. Position-level data is also what allows m⁵C to be linked to a specific isotype rather than to a bulk signal (1).
Bisulfite conversion deaminates unmodified cytidine to uracil, read as thymine, while 5-methylcytidine is protected and read as cytosine. Arraystar applies this chemistry in a tRNA workflow and anchors each protected position to the mature transcript and its structural domain (1,2).
From total RNA to base-resolution m⁵C calls
The workflow runs in one line: RNA sample QC with small-RNA-retaining purification, tRNA processing and bisulfite conversion, library construction and sequencing, then conversion-aware alignment and per-site m⁵C calling.
Bioinformatics for tRNA m⁵C BS-seq
Reads are aligned to a curated tRNA reference, C-to-T conversion signatures are called, and m⁵C stoichiometry is reported per position together with motif, distribution, differential, enrichment and genome-browser results.
Research directions for tRNA m⁵C
Places m⁵C inside anticodon and wobble context to test decoding effects.
Compares m⁵C between the acceptor stem, D-loop, anticodon and TΨC regions.
Contrasts m⁵C occupancy between tRNA isotypes and isoacceptors.
Tests how methyltransferase perturbations shift m⁵C at named positions.
Identifies differentially methylated tRNA positions between groups.
Sample handling and submission for tRNA m⁵C
| Sample Type | Notes |
|---|---|
| Whole blood | Use EDTA tubes only; heparin is not compatible with subsequent analytical procedures. |
| Cultured cells | Cell pellets are preferred to ensure high-quality material for processing. |
| Tissue | Provide fresh or frozen specimens and avoid necrotic material. |
| Total RNA | Maintain an OD 260/280 ratio ≥ 1.8 and RIN ≥ 7 with no visible degradation. Submit at least 10 µg total RNA. |
Submit at least 10 µg total RNA per sample; the amount includes material consumed by sample QC. Questions: support@arraystar.com
Questions we are asked about tRNA m⁵C BS-seq
Key references for tRNA m⁵C
Arraystar tRNA m⁵C BS-seq converts bisulfite chemistry into annotated, quantitative site calls — send us your samples and we will take the project from QC to report.