Disclosed is a method for sequencing a polynucleotide analyte comprising:•a. generating a stream of droplets containing a single nucleotide wherein the order of single nucleotides in the droplet stream corresponds to the sequence of nucleotides in the analyte;•b. introducing into each droplet a plurality of biological probe types each type comprising a different label in an undetectable state and being adapted to capture a different single nucleotide;•c. causing the single nucleotide contained in the droplet to bind to its complementary probe and•d. causing the label to be released from the probe that has bound the nucleotide in a detectable state. The probe is a dumbbell shaped probe comprising fluorescent donor and quencher labels and a single nucleotide gap. After gap repair by a polymerase and a ligase, a restriction enzyme recognition site is cleaved by a restriction enzyme, followed by exonuclease digestion to release the labels.
申请公布号
EP2904109(A1)
申请公布日期
2015.08.12
申请号
EP20130774485
申请日期
2013.10.04
申请人
BASE4 INNOVATION LTD
发明人
FRAYLING, CAMERON ALEXANDER;BALMFORTH, BARNABY;SOARES, BRUNO FLAVIO NOGUEIRA DE SOUSA;ISAAC, THOMAS HENRY;BREINER, BORIS;NATALE, ALESSANDRA;AMASIO, MICHELE