Suspension for determining sequence of genetic materials, method of determining the sequence of genetic materials by using same, and method for high-speed scoring SNPs

A suspension for determining the sequence of genetic materials, wherein suspended in a liquid in a container there is,

• a first coded oligonucleotide group which includes first coded oligonucleotides having one type of base sequence of a predetermined base number which have been coded by molar ratio of labeled material so as to identify the base sequence thereof, so as to cover the base sequences of all the types or specific types for the base number thereof, and which are each provided so as to be sortable as one belonging to this group,
• a second coded oligonucleotide group which includes second coded oligonucleotides having one type of base sequence of a predetermined base number which have been coded by molar ratio of labeled material so as to identify the base sequence thereof, so as to cover the base sequences of all the types or specific types for the base number thereof, and which are each provided so as to be sortable as one belonging to this group,
• wherein said predetermined base number of said respective first coded oligonucleotides and second coded oligonucleotides is at least three bases, and the first coded oligonucleotide group comprises at least 43 types of first coded oligonucleotides obtained by replacing the respective bases of the three bases, and the second coded oligonucleotide group comprises at least 43 types of second coded oligonucleotides obtained by replacing the respective bases of the three bases, and
• an enzyme which differentially bonds the two coded oligonucleotides via a deoxyribose being a sugar of the end nucleotide thereof, only in the case of a situation where the two coded oligonucleotides are hybridized to a single strand genetic material and the end nucleotides of the base sequence thereof are directly adjacent, and the base sequence of said predetermined base number is a constant relation with the base sequence of that genetic material.

A method of determining the sequence of genetic materials using a suspension for determining the sequence of genetic materials, having:

• a conjugation step for introducing and suspending in the suspension, in a container there is,
• a first coded oligonucleotide group which includes first coded oligonucleotides having one type of base sequence of a predetermined base number which have been coded by molar ratio of labeled material so as to identify the base sequence thereof, so as to cover the base sequences of all the types or specific types for the base number thereof, and which are each provided so as to be sortable as one belonging to this group,
• wherein the respective first coded oligonucleotides belonging to said first coded oligonucleotide group comprise an oligonucleotide of one type having a predetermined base sequence and labeled material bonded to said oligonucleotide, and said labeled material includes predetermined kinds thereof at predetermined molar ratios respectively, and the coding is performed by changing the kind and/or the molar ratio of the labeled material,
• a second coded oligonucleotide group which includes second coded oligonucleotides having one type of base sequence of a predetermined base number which have been coded by molar ratio of labeled material so as to identify the bases sequence thereof, so as to cover the base sequences of all the types or specific types for the base number thereof and which are each provided so as to be sortable as one belonging to this group, and
• an enzyme which differentially bonds the two coded oligonucleotides via a deoxyribose being a sugar of the end nucleotide thereof, only in the case of a situation where the two coded oligonucleotides are hybridized to a single strand genetic material and the end nucleotides of the base sequence thereof are directly adjacent, and the base sequence of said predetermined base number is a constant relation with the base sequence of that genetic material, a target genetic material of a single strand of a predetermined base number which is larger than said predetermined base number of the two coded oligonucleotides, for hybridizing said first coded oligonucleotide, and second coded oligonucleotide on said target genetic material;
• a disassociation step for disassociating said target genetic material which has captured the first coded oligonucleotide or the second coded oligonucleotide, into a single stand;
• a sorting step for sorting pairs of the first coded oligonucleotide and the second coded oligonucleotide from in said suspension liquid;
• and
• a determining step for determining the base sequence of the target genetic material based on a combination of the code for identifying the base sequence showing the sorted first coded oligonucleotide, and a code for identifying the base sequence showing the second coded oligonucletide.

A method of determining the sequence of genetic materials using a suspension for determining the sequence of genetic materials, having:

• a conjugation step for introducing and suspending in the suspension, in a container there is,
• a first coded oligonucleotide group which includes first coded oligonucleotides having one type of base sequence of a predetermined base number which have been coded by molar ratio of labeled material so as to identify the base sequence thereof, so as to cover the base sequences of all the types or specific types for the base number thereof, and which are each provided so as to be sortable as one belonging to this group,
• a second coded oligonucleotide group which includes second coded oligonucleotides having one type of base sequence of a predetermined base number which have been coded by molar ratio of labeled material so as to identify type base sequence thereof, so as to cover the base sequences of all the types or specific types for the base number thereof, and which are each provided so as to be sortable as one belonging to this group,
• wherein said predetermined base number of said respective first coded oligonucleotides and second coded oligonucleotides is at least three bases, and the first coded oligonucleotides group comprises at least 43 types of first coded oligonucleotides obtained by replacing the respective bases of the three bases, and the second coded oligonucleotide group comprises at least 43 types of second coded oligonucleotides obtained by replacing the respective bases of the three bases and
• an enzyme which differentially bonds the two coded oligonucleotides via a deoxyribose being a sugar of the end nucleotide thereof, only in the case of a situation where the two coded oligonucleotides are hybridized to a single strand genetic material and the end nucleotides of the base sequence thereof are directly adjacent, and the base sequence of said predetermined base number is a constant relation with the base sequence of that genetic material, a target genetic material of a single strand of a predetermined base number which is larger than said predetermined base number of the two coded oligonucleotides, for hybridizing said first coded oligonucleotide, and second coded oligonucleotide on said target genetic material;
• a disassociation step for disassociating said target genetic material which has captured the first coded oligonucleotide or the second coded oligonucleotide, into a single strand;
• a sorting step for sorting pairs of the first coded oligonucleotide and the second coded oligonucleotide from in said suspension liquid;
• and
• a determining step for determining the base sequence of the target genetic material based on a combination of the code for identifying the base sequence showing the sorted first coded oligonucleotide, and a code for identifying the base sequence showing the second coded oligonucleotide.

A method for high-speed scoring SNPs using a suspension for determining the sequence of genetic materials having:

• a conjugation step for preparing two approximately identical suspensions, in each container there is,
• a first coded oligonucleotide group which includes first coded oligonucleotides having one type of base sequence of a predetermined base number which have been coded by molar ratio of labeled material so as to identify the base sequence thereof, so as to cover the base sequences of all the types or specific types for the base number thereof, and which are each provided so as to be sortable as one belonging to this group,
• a second coded oligonucleotide group which includes second coded oligonucleotides having one type of base sequence of a predetermined base number which have been coded by molar ratio of labeled material so as to identify the base sequence thereof, so as to cover the base sequences of all the types dr specific types for the base number thereof, and which are each provided so as to be sortable as one belonging to this group,
• wherein said predetermined base number of said respective first coded oligonucleotides and second coded oligonucleotides is at least three bases, and the first coded oligonucleotide croup comprises at least 43 types of first coded oligonucleotides obtained by replacing the respective bases of the three bases, and the second coded oligonucleotide group, comprises at least 43 types of second coded oligonucleotides obtained by replacing the respective bases of the three bases, and
• an enzyme which differentially bonds the two coded oligonucleotides via a deoxyribose being a sugar of the end nucleotide thereof, only in the case of a situation where the two coded oligonucleotides are hybridized to a single strand genetic material and the end nucleotides of the base sequence thereof are directly adjacent, and the base sequence of said predetermined base number is a constant relation with the base sequence of that genetic material, and introducing and suspending in one thereof a first target genetic material of a single strand of a predetermined base number obtained by extracting from a first specimen, and introducing and suspending in the other thereof a second target genetic material of a single strand of a predetermined base number obtained by extracting from a second specimen, and hybridizing said first coded oligonucleotide, and second coded oligonucleotide on respective target materials;
• a disassociation step for again disassociating said target genetic material which has captured each of the first coded oligonucleotide or the second coded oligonucleotide, into a single strand;
• a sorting step for sorting the first coded oligonucleotide and the second coded oligonucleotide from in said suspension liquid;
• and
• a determining step for determining the base sequence of the target genetic material based on each of a combination of the code for identifying the base sequence showing the sorted first coded oligonucleotide, and a code for identifying the base sequence showing the second coded oligonucleotide;
• and
• an identification step for performing identification of single nucleotide polymorphisms by comparing base sequences determined for said first target genetic material and base sequences determined for said second target genetic material.