Abstract: The embodiments of the application provide a base-calling method and device, electronic equipment and a storage medium, and belongs to the technical field of recognition. The base-calling method comprises: acquiring original light intensity data of original base channels; performing crosstalk correction on the original light intensity data to obtain first corrected light intensity data; performing quantile normalization on the first corrected light intensity data to obtain initial light intensity data; performing phase correction on the initial light intensity data to obtain second corrected light intensity data; performing mean normalization on the second corrected light intensity data to obtain target light intensity data; performing intensity comparison and screening on the original base channels according to the target light intensity data to obtain a target base; and splicing multiple target bases to obtain a target base sequence. The embodiments of the application can improve the base-calling accuracy.

Abstract: A gene sequencing method includes: hybridizing a sequencing primer and a molecule to be detected to form a template strand and a primer strand; linking a first nucleotide analog to the primer strand, wherein the first nucleotide analog has a blocking group; performing base pairing on a second nucleotide analog and the nucleic acid molecule to be detected, the second nucleotide analog forming a complex with the nucleic acid molecule to be detected and the first nucleotide analog under the action of metal ions and a polymerase, wherein the second nucleotide analog has a marker; detecting the marker, and identifying a base; and removing the blocking group and the second nucleotide analog, and performing a next cycle of sequencing. The gene sequencing method can completely remove blocking groups and markers without leaving synthetic scars, so that the sequencing length can be increased and the sequencing cost can be reduced.

Abstract: The present invention relates to a nucleoside analog having the structure of the following formula, a method for preparing the nucleoside analog, and an application of the nucleoside analog in nucleic acid sequencing, etc. wherein L1, L2, and L3 are each independently a covalent bond or a covalently linked group; B is a base or a base derivative; R1 is —OH or a phosphate group; R2 is H or a cleavable group; R3 is a detectable group or a targeting group; R5 is a polymerase reaction inhibitory group; R4 is H or —OR6, wherein R6 is H or a cleavable group; and C is a cleavable group or a cleavable bond.

Abstract: The present invention relates to a nucleoside analog having the structure of the following formula, a method for preparing the nucleoside analog, and an application of the nucleoside analog in nucleic acid sequencing, etc. wherein L1, L2, and L3 are each independently a covalent bond or a covalently linked group; B is a base or a base derivative; R1 is —OH or a phosphate group; R2 is H or a cleavable group; R3 is a detectable group or a targeting group; R5 is a polymerase reaction inhibitory group; R4 is H or —OR6, wherein R6 is H or a cleavable group; and C is a cleavable group or a cleavable bond.

Abstract: Nucleoside or nucleotide analog compounds having the structure shown below, a method for preparing them, and applications in nucleic acid sequencing are disclosed. The compounds have formula (I): wherein L1, L2, and L3 are each independently a covalent bond or a covalently linked group; B is a base or a base derivative selected from purines, pyrimidines, or analogs thereof; R1 is —OH, a phosphate group, or a nucleotide; R2 as H or a cleavable group; R3 is a detectable group or a targeting group; R5 is an inhibitory group; R4 is H, —NH2, or —OR6, wherein R6 is H or a cleavable group; and C is a cleavable group or a cleavable bond.

Abstract: Disclosed are a method for ligating nucleic acid fragments, a method for constructing a sequencing library, and the use thereof. The method for ligating nucleic acid fragments comprises ligating a first nucleic acid fragment and a second nucleic acid fragment in a mixed enzyme reaction system comprising a DNA ligase and a RNA ligase, wherein the first nucleic acid fragment is double-stranded DNA. The ligation efficiency of the method for ligating nucleic acid fragments can reach 85% or more, and by using this ligation method to construct a sequencing library, the yield of the effective library is significantly improved.

Abstract: A chip and a manufacturing method for the chip, comprising a weak passivation treatment following a fixing treatment. The weak passivation treatment comprises putting a weak passivation reaction solution containing a catalyst into contact with a chip for the fixing treatment, so as to promote a DNA or protein to bond with a substrate surface, thus allowing the DNA or protein to be fully fixed on the substrate surface. Moreover, on such basis, a small molecule or protein is added for manufacturing the chip.

Abstract: The present disclosure discloses a method, a device and a system for detecting chromosomal aneuploidy. The method comprises: sequencing at least a portion of a nucleic acid in a sample under test to obtain a sequencing result including reads; aligning the reads to a first reference sequence to obtain an alignment result including specific chromosomes to which the reads are mapped; determining, for a first chromosome, the amount of reads mapped to the first chromosome based on the alignment result; and comparing the number of the reads mapped to the first chromosome with the amount of reads in a negative control mapped to the first chromosome to determine the number of the first chromosome. When the method is employed to detect chromosomal aneuploidy, the detection results acquired have relatively high sensitivity and accuracy.

Abstract: Nucleoside or nucleotide analog compounds having the structure shown below, a method for preparing them, and applications in nucleic acid sequencing are disclosed. The compounds have formula (I): wherein L1, L2, and L3 are each independently a covalent bond or a covalently linked group; B is a base or a base derivative selected from purines, pyrimidines, or analogs thereof; R1 is —OH, a phosphate group, or a nucleotide; R2 as H or a cleavable group; R3 is a detectable group or a targeting group; R5 is an inhibitory group; R4 is H, —NH2, or —OR6, wherein R6 is H or a cleavable group; and C is a cleavable group or a cleavable bond.

Abstract: The present disclosure discloses a total internal reflection fluorescence imaging system and a sequencing device. The total internal reflection fluorescence imaging system includes a first imaging system and a second imaging system. The first imaging system includes N separate laser light paths. The second imaging system determines the changes of the height of the sample by means of the differences of the positions at which the second receiving device acquires the second images, and performs automatic compensation according to the changes of the height of the sample, so as to ensure that the sample is always at a focused position. The embodiment can perform light path adjustment of the N separate laser light paths respectively by means of the second imaging system, so as to realize that the N separate laser light paths have the same penetration depth. Therefore no further light path adjustment is required when switching between different light sources, thus saving the time consuming for light path adjustment.

Abstract: The present disclosure discloses a total internal reflection fluorescence imaging system and a sequencing device. The total internal reflection fluorescence imaging system includes a first imaging system and a second imaging system. The first imaging system includes N separate laser light paths. The second imaging system determines the changes of the height of the sample by means of the differences of the positions at which the second receiving device acquires the second images, and performs automatic compensation according to the changes of the height of the sample, so as to ensure that the sample is always at a focused position. The embodiment can perform light path adjustment of the N separate laser light paths respectively by means of the second imaging system, so as to realize that the N separate laser light paths have the same penetration depth. Therefore no further light path adjustment is required when switching between different light sources, thus saving the time consuming for light path adjustment.