smMIP
smMIP
The single-molecule molecular inversion probes (smMIP) method uses single-molecule tagging and molecular inversion probes to detect and quantify genetic variations occurring at very low frequencies. In this method, probes are used to detect targets in genomic DNA. After the probed targets are copied, exonuclease digestion leaves the target with a tag, which undergoes PCR amplification and sequencing. Sequencing allows for high-resolution sequence reads of targets, while greater depth allows for better alignment for every unique molecular tag.
Pros:
- Detection of low-frequency targets
- Can perform single-cell sequencing or sequencing for samples with very limited starting material
Cons:
- PCR amplification errors
- PCR biases can underrepresent GC-rich templates
- Targets smaller than 500 bp are preferentially amplified by polymerases during PCR