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Does scFAST-seq technology achieve complete transcript coverage? Can all genes in a single cell be fully covered?
2026-06-23
Answer:
scFAST-seq achieves population-level full-length coverage, but not single-molecule full-length sequencing.
- Technology principle:
- Third-generation sequencing: directly sequences single full-length molecules
- scFAST-seq (NGS-based): uses short reads + random priming
- Third-generation sequencing: directly sequences single full-length molecules
- Coverage mechanism:
- Reverse transcription initiates at random positions along RNA molecules
- For highly expressed genes, multiple fragments collectively reconstruct near full-length coverage
- This represents aggregate coverage, not continuous reads from a single molecule
- Reverse transcription initiates at random positions along RNA molecules
- Single-cell limitation:
- Not all genes in a single cell can achieve full-length coverage
- Coverage depends strongly on gene expression level
- Not all genes in a single cell can achieve full-length coverage
- Low-expression genes:
If only a few reads are detected (e.g., 1 read per gene), full-length reconstruction is not possible.
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