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US 12,723,276 B2Granted
Method to calibrate nucleic acid library seeding efficiency in flowcells
AssigneeIllumina, Inc.
InventorsYir-Shyuan Wu, Filiz Gorpe-Yasar, Jeff Fisher, Jeff Brodin
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Abstract
The disclosure provides methods to calibrate polynucleotide seeding efficiency in flow cells.
Claims21
A method to evaluate the seeding efficiency of a flow cell with polynucleotides, comprising:
- seeding a flow cell with a known concentration of polynucleotides for at least 1 minute, and
- collecting the supernatant;
- quantifying the polynucleotides in the supernatant by using step (a) or (b):
- (a) amplifying the polynucleotides in the supernatant using qPCR and/or droplet PCR;
- or
- (b) reseeding the supernatant using a second flow cell and counting clusters generated after bridge amplification of the polynucleotides;
- and
- (c) determining seeding efficiency of the flow cell by comparing the number of polynucleotides quantified in the supernatant versus the number of polynucleotides used to seed the flow cell.
5 more elements
Dependencies are inferred from the claim text.