	US 11,795,451 B2
	Primer for next generation sequencer and a method for producing the same, a DNA library obtained through the use of a primer for next generation sequencer and a method for producing the same, and a DNA analyzing method using a DNA library
Hiroyuki Enoki, Hamamatsu (JP); Yoshie Takeuchi, Hamamatsu (JP); and Minoru Inamori, Kariya (JP)
Assigned to TOYOTA JIDOSHA KABUSHIKI KAISHA, Toyota (JP)
Appl. No. 16/769,859
Filed by TOYOTA JIDOSHA KABUSHIKI KAISHA, Toyota (JP)
PCT Filed Dec. 21, 2018, PCT No. PCT/JP2018/047136 § 371(c)(1), (2) Date Jun. 4, 2020, PCT Pub. No. WO2019/131470, PCT Pub. Date Jul. 4, 2019.
Claims priority of application No. 2017-247826 (JP), filed on Dec. 25, 2017.
Prior Publication US 2021/0171941 A1, Jun. 10, 2021
Int. Cl. G16B 30/00 (2019.01); G16B 40/00 (2019.01); G16B 25/20 (2019.01); C12N 15/10 (2006.01); C12Q 1/6806 (2018.01)

CPC C12N 15/1093 (2013.01) [C12Q 1/6806 (2013.01); G16B 25/20 (2019.02); G16B 30/00 (2019.02); G16B 40/00 (2019.02); C12Q 2537/165 (2013.01)]

5 Claims

1. A method for producing a primer comprising a nucleotide sequence of 5′-CAAGCAGAAGACGGCATACGAGAT (SEQ ID NO: 1)-N_{5 to 15}-GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAG (SEQ ID NO: 2)-3′, wherein N_{5 to 15}represents an index sequence of 5 to 15 nucleotides, the method comprising:

(i) calculating for the primer a putative number of reads for multiple index sequences based on a nucleotide sequence of the index sequence in accordance with an estimation formula that designates the putative number of reads as a purpose variable and a type of a nucleotide for each of the nucleotides in the index sequence as an explanatory variable, to select a nucleotide sequence as a nucleotide sequence of the index sequence from index sequences that have a putative number of reads that exceed a given level; and

(ii) synthesizing a nucleotide sequence comprising the nucleotide sequence selected in (i), wherein the synthesizing nucleotide sequence is the primer comprising the nucleotide sequence of SEQ ID NO: 1 and SEQ ID NO: 2 and the selected index sequence.