Retrieval of DNA sequences present at an extremely low frequency.
← Research streamA method using stringent hybridization to an immobilized probe followed by dissociation and amplification, repeated twice, retrieved target sequences with one or two point mutations present at very low frequencies (1 in 466×10^3 and 1 in 28×10^3 molecules). T…
Study snapshot
Abstract
We describe a method for retrieving sequences with one or two point mutations of a given target sequence, which are present in a DNA population at a frequency of 1 in 466 x 10(3) and 1 in 28 x 10(3) molecules, respectively. By stringent hybridization to a stable, chemically immobilized probe, a large excess of unrelated fragments is removed, and the bound sequences are dissociated and amplified. By repeating the hybridization-amplification cycles twice, we achieved an estimated enrichment of 404,000-fold and 1612-fold, respectively, which was confirmed by cloning the resultant products and sequencing 35 clones. This procedure can be applied to retrieve mutated sequences that exist at an extremely low frequency in a DNA population.
AI evidence extraction
Structured fields below were generated from the paper’s abstract and metadata to make the evidence easier to inspect. Automated extraction may be incomplete or incorrect; verify important details against the original paper.
Main findings
A method using stringent hybridization to an immobilized probe followed by dissociation and amplification, repeated twice, retrieved target sequences with one or two point mutations present at very low frequencies (1 in 466×10^3 and 1 in 28×10^3 molecules). The authors report estimated enrichment of 404,000-fold and 1,612-fold, confirmed by cloning and sequencing 35 clones.
Outcomes measured
- Retrieval/enrichment of rare DNA sequences with point mutations from a DNA population
- Estimated enrichment fold-change after hybridization-amplification cycles
- Confirmation by cloning and sequencing
Limitations
- No EMF exposure or health-related outcomes described in the abstract
- Sample size for sequencing confirmation is limited to 35 clones (as stated)
- Frequencies/enrichment are reported as estimates; details of estimation not provided in the abstract
View raw extracted JSON
{
"study_type": "other",
"exposure": {
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"outcomes": [
"Retrieval/enrichment of rare DNA sequences with point mutations from a DNA population",
"Estimated enrichment fold-change after hybridization-amplification cycles",
"Confirmation by cloning and sequencing"
],
"main_findings": "A method using stringent hybridization to an immobilized probe followed by dissociation and amplification, repeated twice, retrieved target sequences with one or two point mutations present at very low frequencies (1 in 466×10^3 and 1 in 28×10^3 molecules). The authors report estimated enrichment of 404,000-fold and 1,612-fold, confirmed by cloning and sequencing 35 clones.",
"effect_direction": "unclear",
"limitations": [
"No EMF exposure or health-related outcomes described in the abstract",
"Sample size for sequencing confirmation is limited to 35 clones (as stated)",
"Frequencies/enrichment are reported as estimates; details of estimation not provided in the abstract"
],
"evidence_strength": "insufficient",
"confidence": 0.78,
"peer_reviewed_likely": "yes",
"keywords": [
"DNA sequence retrieval",
"rare mutations",
"point mutations",
"hybridization",
"immobilized probe",
"PCR amplification",
"enrichment",
"cloning",
"sequencing"
],
"suggested_hubs": []
}AI-extracted fields are generated from the abstract and metadata and may be incomplete or incorrect. This material is for informational purposes and is not medical advice.
Identifiers and source
DOI: 10.2144/97236rr02
PubMed: 9421644