Methods of specifically labeling nucleic acids using CRISPR/Cas
US10640810B2 · kind B2 · utility
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Key dates
| Filing date | Oct 18, 2017 |
| Grant date | May 5, 2020 |
| Priority date | — |
| Expiry date | Oct 20, 2037 |
Classification
- Technology area (CPC G)Physics
- CPC primaryG01N2021/6439
- WIPO fieldMeasurement
- WIPO sectorInstruments
Abstract
Provided herein are methods of detecting a target nucleic acid sequence. In one embodiment, the method includes contacting genomic DNA with a guide RNA having a portion complementary to the target sequence in the genomic DNA and with Cas9 nickase to produce a single-strand break in the genomic DNA at a specific location adjacent to the target sequence. The method further includes contacting the nicked DNA with a polymerase and fluorescently labeled nucleotide. The fluorescently labeled nucleotide is incorporated into the nicked DNA at the specific location and the target nucleic acid sequence is detected via fluorescent label.
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