A polynucleotide sequencing method includes a wash step that employs a composition including a polymerase. The composition may also include a plurality of nucleotides. The composition may be configured to prevent the polymerase from incorporating one of the plurality of nucleotides into a copy polynucleotide strand. The composition may be substantially free of Mg.sup.2+.

The present invention relates to a method of spatially barcoding a given location on a substrate, and further to spatially barcoding detection probes present in a sample such as a biological tissue specimen for the purposes of analysing molecular features present in the tissue. Such analysis may include: i) the spatial expression of one or more biological molecules, specifically; ii) the spatial analysis of the transcriptome and/or iii) the spatial analysis of the proteome, including post-translational protein modifications. The invention further relates to various component products for performing such methods that include reagents kits, instrumentation and software.

The present invention relates to a method of spatially barcoding a given location on a substrate, and further to spatially barcoding detection probes present in a sample such as a biological tissue specimen for the purposes of analysing molecular features present in the tissue. Such analysis may include: i) the spatial expression of one or more biological molecules, specifically; ii) the spatial analysis of the transcriptome and/or iii) the spatial analysis of the proteome, including post-translational protein modifications. The invention further relates to various component products for performing such methods that include reagents kits, instrumentation and software.

An apparatus for preparing nucleic acid sequences using an enzyme, including a reactor, a plurality of nucleotide material bottles, a deblocking material bottle, and a liquid delivering device. The reactor includes a reaction substrate having a pretreated surface. Each of the nucleotide material bottles is adapted to contain a first reaction solution, and the first reaction solution includes a reaction enzyme and a nucleotide having a terminal protecting group. The deblocking material bottle is adapted to contain a deblocking solution. The liquid delivering device is connected to the reactor, the nucleotide material bottles and the deblocking material bottle. The reaction enzyme is adapted to dispose the nucleotide having the terminal protecting group on the pretreated surface. The reactor has an operating temperature of 45° C.-105° C. A method for preparing nucleic acid sequences using an enzyme of the invention is provided.

An apparatus for preparing nucleic acid sequences using an enzyme, including a reactor, a plurality of nucleotide material bottles, a deblocking material bottle, and a liquid delivering device. The reactor includes a reaction substrate having a pretreated surface. Each of the nucleotide material bottles is adapted to contain a first reaction solution, and the first reaction solution includes a reaction enzyme and a nucleotide having a terminal protecting group. The deblocking material bottle is adapted to contain a deblocking solution. The liquid delivering device is connected to the reactor, the nucleotide material bottles and the deblocking material bottle. The reaction enzyme is adapted to dispose the nucleotide having the terminal protecting group on the pretreated surface. The reactor has an operating temperature of 45° C.-105° C. A method for preparing nucleic acid sequences using an enzyme of the invention is provided.

Nucleic acid memory strands encoding digital data using a sequence of homopolymer tracts of repeated nucleotides provides a cheaper and faster alternative to conventional digital DNA storage techniques. The use of homopolymer tracts allows for lower fidelity, high throughput sequencing techniques such as nanopore sequencing to read data encoded in the memory strands. Specialized synthesis techniques allow for synthesis of long memory strands capable of encoding large volumes of data despite the reduced data density afforded by homopolymer tracts as compared to conventional single nucleotide sequences.

Nucleic acid memory strands encoding digital data using a sequence of homopolymer tracts of repeated nucleotides provides a cheaper and faster alternative to conventional digital DNA storage techniques. The use of homopolymer tracts allows for lower fidelity, high throughput sequencing techniques such as nanopore sequencing to read data encoded in the memory strands. Specialized synthesis techniques allow for synthesis of long memory strands capable of encoding large volumes of data despite the reduced data density afforded by homopolymer tracts as compared to conventional single nucleotide sequences.

An oligonucleotide containing a blocker, relating to the field of target sequence hybridization capture and the design and synthesis of universal blocking oligonucleotides. Applying an oligonucleotide that has a blocking function or a combination thereof not only has a good blocking effect on an linker sequence during the capture of a target sequence in a single sample, but also reduces non-specific capture and improves the capture efficiency. In particular, the oligonucleotide may effectively block linker sequences at both ends of a target sequence in a plurality of samples and improve the target sequence capture efficiency.

An oligonucleotide containing a blocker, relating to the field of target sequence hybridization capture and the design and synthesis of universal blocking oligonucleotides. Applying an oligonucleotide that has a blocking function or a combination thereof not only has a good blocking effect on an linker sequence during the capture of a target sequence in a single sample, but also reduces non-specific capture and improves the capture efficiency. In particular, the oligonucleotide may effectively block linker sequences at both ends of a target sequence in a plurality of samples and improve the target sequence capture efficiency.

Provided is a method for synchronously sequencing a sense strand and an antisense strand of an insert DNA, including: performing two rounds of rolling circle amplification and multiple displacement amplification to obtain a DNA nano ball template including a read1 strand sequencing template and a read2 strand sequencing template; and hybridizing the read1 strand sequencing template and the read2 strand sequencing template with read1 strand sequencing primers and read2 strand sequencing primers, respectively, and simultaneously performing read1 strand sequencing and read2 strand sequencing to obtain sequences of the sense strand and the antisense strand of the insert DNA. The method of the present disclosure can perform the sequencing from both ends of the insert DNA, significantly saving the time and costs for sequencing, and increasing the sequencing throughput.