Method for maintaining nanopore sequencing speed
AssigneeBgi Shenzhen
InventorsZhouxiang JI, Jingxiong LU, Rongrong ZHENG, Wenwei ZHANG, Yuliang DONG, Yuxiang LI, Tao ZENG, Xun XU
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Abstract
Provided in the present disclosure is a method for sequencing a double-stranded target polynucleotide. The method can keep ATP at a relatively constant concentration during sequencing, so that the sequencing rate can be better kept stable and unchanged. Further provided in the present disclosure is a kit for sequencing a double-stranded target polynucleotide, the kit including a transmembrane pore in the membrane, a helicase, an ATP-generating enzyme and an ATP-generating substrate.
Claims19
A method for sequencing a double-stranded target polynucleotide, comprising:
- (a) providing a double-stranded target polynucleotide, a membrane containing a transmembrane pore, a helicase, an ATP-generating enzyme, and an ATP-generating substrate, wherein the ATP-generating substrate is capable of reacting with ADP to generate ATP under the catalysis of the ATP-generating enzyme;
- (b) contacting the double-stranded target polynucleotide with the transmembrane pore, the helicase, the ATP-generating enzyme, and the ATP-generating substrate, wherein double strands of the double-stranded target polynucleotide are separated by the helicase to form a single-stranded target polynucleotide, and wherein the helicase enables the single-stranded target polynucleotide to move in the transmembrane pore, allowing a portion of nucleotides in the single-stranded target polynucleotide to interact with the transmembrane pore;
- and
- (c) measuring current that passes through the transmembrane pore during each interaction to determine the sequence of the double-stranded target polynucleotide.
A kit, comprising a membrane containing a transmembrane pore, a helicase, an ATP-generating enzyme, and an ATP-generating substrate.
Dependencies are inferred from the claim text.