Conjoined synthetic polynucleotides
AssigneeGarvan Institute of Medical Research
InventorTimothy Mercer
Life of this patent
Estimating the term…
Abstract
This disclosure includes compositions comprising different synthetic polynucleotides that can be used in determining whether a given gene mutation (e.g., a cancer-associated mutation) is present in a target polynucleotide and/or determining an abundance of a target polynucleotide (e.g., by amplifying and sequencing the different synthetic polynucleotides and the target polynucleotide). In some aspects, the synthetic polynucleotides each comprise a barcode, which can be used to differentiate identical synthetic polynucleotides and differentiate the synthetic polynucleotides and the target polynucleotide.
Claims21
-19. (canceled)
A composition for use in determining an abundance of one or more target polynucleotides in a sample, the composition comprising, for each target polynucleotide of a plurality of different target polynucleotides:
- multiple synthetic polynucleotides designed to be amplified with the target polynucleotide using a single pair of primers, each of the multiple synthetic polynucleotides comprising:
- (i) a first primer binding site that is also present in the target polynucleotide;
- (ii) a barcode that can be used to distinguish among otherwise identical synthetic polynucleotides;
- and
- (iii) a reverse complement of a second primer binding site, wherein the reverse complement of the second primer binding site is also present in the target polynucleotide,
- wherein the plurality of different target polynucleotides comprises polynucleotides of each of a serine/threonine kinase B-Raf (BRAF) gene, a Kirsten rat sarcoma viral oncogene homolog (KRAS) gene, an epidermal growth factor receptor (EGFR) gene, a fibroblast growth factor receptor 3 (FGFR3) gene, a phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA) gene, a KIT proto-oncogene receptor tyrosine kinase (KIT) gene, and an N-ras proto-oncogene (NRAS) gene.
A composition for use in determining an abundance of one or more target polynucleotides in a sample, the composition comprising:
- (a) for each target polynucleotide of a plurality of different target polynucleotides:
- multiple synthetic polynucleotides designed to be amplified with the target polynucleotide using a single pair of primers, each of the multiple synthetic polynucleotide comprising:
- (i) a first primer binding site that is also present in the target polynucleotide;
- (ii) a 10 contiguous nucleotide barcode that can be used to distinguish among otherwise identical synthetic polynucleotides;
- and
- (iii) a reverse complement of a second primer binding site, wherein the reverse complement of the second primer binding site is also present in the target polynucleotide,
- (b) a sample comprising one or more of the target polynucleotides of the plurality of different target polynucleotides, wherein the plurality of different target polynucleotides comprises polynucleotides of each of a BRAF gene, a KRAS gene, an EGFR gene, an FGFR3 gene, a PIK3CA gene, a KIT gene, and an NRAS gene.
Dependencies are inferred from the claim text.