Abstract: The present invention provides a method for amplifying a sequence adjacent to a specific sequence, comprising the steps of: annealing a first forward primer to the specific sequence to synthesize a complementary strand; sequentially polymerically adding a first deoxynucleotide and a second deoxynucleotide to a 3?-end of the complementary strand; annealing a first reverse primer to a binding site between the 3?-end of the complementary strand and a polydeoxynucleotide strand composed of the first deoxynucleotide to synthesize a double-stranded DNA; performing a PCR with the double-stranded DNA as a template by using a second forward primer complementary to the specific sequence and a first reverse primer; and further performing a PCR by using a third forward primer complementary to the specific sequence and a second reverse primer.

Abstract: An object of the present invention is to provide a method of conveniently producing a genetically modified non-human mammal with high efficiency using a CRISPR-Cas9 system and particularly a production method whereby gene knock-in can be achieved with high efficiency regardless of the gene size. The method of producing a genetically modified non-human mammal comprises introducing a Cas9 protein, a crRNA fragment comprising a nucleotide sequence complementary to a target DNA region, and a tracrRNA fragment into a non-human mammalian oocyte to genetically modify the target DNA.

Abstract: The present invention relates to a method of editing a filamentous fungal genome by direct introduction of a genome editing protein molecule or complex. The method includes three modes. In the first mode, a genome editing protein molecule or complex for a target gene is directly introduced into a cell of an Aspergillus fungus, to edit a gene in the Aspergillus fungal genome. In the second mode, a genome editing protein molecule or complex for a target region in a filamentous fungal genome, and a desired DNA fragment, are directly introduced into a filamentous fungal cell, to knock-in the DNA fragment to a desired target site in the filamentous fungal genome. In the third mode, genome editing protein molecules or complexes for plural target genes are directly introduced into a filamentous fungal cell, to carry out simultaneous editing of the plural genes in the filamentous fungal genome.

Abstract: An object of the present invention is to provide a method of conveniently producing a genetically modified non-human mammal with high efficiency using a CRISPR-Cas9 system and particularly a production method whereby gene knock-in can be achieved with high efficiency regardless of the gene size. The method of producing a genetically modified non-human mammal comprises introducing a Cas9 protein, a crRNA fragment comprising a nucleotide sequence complementary to a target DNA region, and a tracrRNA fragment into a non-human mammalian oocyte to genetically modify the target DNA.

Abstract: In a clustering apparatus comprising an input unit (1) supplied with a dataset including a plurality of samples, a data processing unit (4) for processing the samples to classify each sample into a class, and an output unit (3) for producing a processing result representative of classification, a parameter memory (51) in a memory unit (5) memorizes a target parameter obtained from past experiment. A parameter estimating section (24) of the data processing unit estimates a clustering parameter by the use of the target parameter memorized in the parameter memory. An unidentifiable sample detecting section (25) of the data processing unit detects a sample as an unidentifiable sample if posterior probabilities calculated for the sample by a probability density function produced by the clustering parameter estimated by the parameter estimating section are smaller than a predetermined value.