Patent classifications
C12Q2525/173
METHOD AND KIT FOR NON-SPECIFIC AMPLIFICATION OF NATURAL SHORT-FRAGMENT NUCLEIC ACID
The present invention relates to a method for a non-specific amplification of a natural short-fragment nucleic acid, comprising the following steps: (1) performing end repair on the natural short-fragment nucleic acid to obtain an end-repaired nucleic acid; (2) connecting the end-repaired nucleic acid to a double-stranded linker to obtain a ligation product, in which each strand of the double-stranded linker contains only three bases; (3) performing PCR amplification on the ligation product using a PCR primer labeled with deoxyuridine to obtain a PCR product, in which the PCR primer is completely or partially complementary to a strand of the double-stranded linker and contains only three bases; and (4) digesting the PCR product by using an enzyme having a deoxyuridine cleavage function, followed by digesting the PCR product by using an enzyme with both 5′.fwdarw.3′ polymerase activity and 3′.fwdarw.5′ exonuclease activity in the presence of a deoxynucleotide solution to obtain a non-specific amplification product of the natural short-fragment nucleic acid. The deoxynucleotide solution only contains the complementary base of the base lacking in the primer. The present invention also relates to a kit for implementing the aforementioned method.
METHOD AND KIT FOR NON-SPECIFIC AMPLIFICATION OF NATURAL SHORT-FRAGMENT NUCLEIC ACID
The present invention relates to a method for a non-specific amplification of a natural short-fragment nucleic acid, comprising the following steps: (1) performing end repair on the natural short-fragment nucleic acid to obtain an end-repaired nucleic acid; (2) connecting the end-repaired nucleic acid to a double-stranded linker to obtain a ligation product, in which each strand of the double-stranded linker contains only three bases; (3) performing PCR amplification on the ligation product using a PCR primer labeled with deoxyuridine to obtain a PCR product, in which the PCR primer is completely or partially complementary to a strand of the double-stranded linker and contains only three bases; and (4) digesting the PCR product by using an enzyme having a deoxyuridine cleavage function, followed by digesting the PCR product by using an enzyme with both 5′.fwdarw.3′ polymerase activity and 3′.fwdarw.5′ exonuclease activity in the presence of a deoxynucleotide solution to obtain a non-specific amplification product of the natural short-fragment nucleic acid. The deoxynucleotide solution only contains the complementary base of the base lacking in the primer. The present invention also relates to a kit for implementing the aforementioned method.
DETERMINISTIC BARCODING FOR SPATIAL OMICS SEQUENCING
Provided herein, in some embodiments, are compositions and methods for producing a molecular expression map of a biological sample using Deterministic Barcoding in Tissue for spatial omics sequencing (DBiT-seq).
DETERMINISTIC BARCODING FOR SPATIAL OMICS SEQUENCING
Provided herein, in some embodiments, are compositions and methods for producing a molecular expression map of a biological sample using Deterministic Barcoding in Tissue for spatial omics sequencing (DBiT-seq).
METHOD FOR OBTAINING SPATIAL AND SEQUENCING INFORMATION OF M-RNA FROM TISSUE
The invention is directed to a method to obtain the spatial location and sequence information of an m-RNA target sequence on a tissue sample comprising providing a solid surface, attaching anchor molecules, binding scaffolding molecules, incorporating adenine, guanine, cytosine and thymine, incorporating thymine to the anchor molecules, removing the scaffolding, providing a tissue sample, reverser transcrining to create c-DNA, removing the c-DNA and obtaining the sequence information of the c-DNA.
METHOD FOR OBTAINING SPATIAL AND SEQUENCING INFORMATION OF M-RNA FROM TISSUE
The invention is directed to a method to obtain the spatial location and sequence information of an m-RNA target sequence on a tissue sample comprising providing a solid surface, attaching anchor molecules, binding scaffolding molecules, incorporating adenine, guanine, cytosine and thymine, incorporating thymine to the anchor molecules, removing the scaffolding, providing a tissue sample, reverser transcrining to create c-DNA, removing the c-DNA and obtaining the sequence information of the c-DNA.
METHODS AND COMPOSITIONS FOR COMBINATORIAL BARCODING
The present disclosure provide compositions, methods and kits for generating a set of combinatorial barcodes, and uses thereof for barcoding samples such as single cells or genomic DNA fragments. Some embodiments disclosed herein provide compositions comprising a set of component barcodes for producing a set of combinatorial barcodes. The set of component barcodes can comprise, for example, n×m unique component barcodes, wherein n and m are integers, each of the component barcodes comprises: one of n unique barcode subunit sequences; and one or two linker sequences or the complements thereof, wherein the component barcodes are configured to connect to each other through the one or two linker sequences or the complements thereof to produce a set of combinatorial barcodes.
METHODS AND COMPOSITIONS FOR COMBINATORIAL BARCODING
The present disclosure provide compositions, methods and kits for generating a set of combinatorial barcodes, and uses thereof for barcoding samples such as single cells or genomic DNA fragments. Some embodiments disclosed herein provide compositions comprising a set of component barcodes for producing a set of combinatorial barcodes. The set of component barcodes can comprise, for example, n×m unique component barcodes, wherein n and m are integers, each of the component barcodes comprises: one of n unique barcode subunit sequences; and one or two linker sequences or the complements thereof, wherein the component barcodes are configured to connect to each other through the one or two linker sequences or the complements thereof to produce a set of combinatorial barcodes.
Methods of producing amplified double stranded deoxyribonucleic acids and compositions and kits for use therein
Methods of producing an amplified double stranded deoxyribonucleic acid (dsDNA) from a nucleic acid sample are provided. Aspects of the methods include amplifying using a single product nucleic acid primer and a template switch oligonucleotide to produce an amplified dsDNA product. Compositions and kits for use in performing the methods are also provided.
Methods of producing amplified double stranded deoxyribonucleic acids and compositions and kits for use therein
Methods of producing an amplified double stranded deoxyribonucleic acid (dsDNA) from a nucleic acid sample are provided. Aspects of the methods include amplifying using a single product nucleic acid primer and a template switch oligonucleotide to produce an amplified dsDNA product. Compositions and kits for use in performing the methods are also provided.