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Tlhahlobo ea Mokhatlo oa Genome ka Bophara
Sepheo sa Lithuto tsa Mokhatlo oa Genome-Wide (GWAS) ke ho khetholla mefuta e fapaneng ea liphatsa tsa lefutso (genotypes) e amanang le litšobotsi tse itseng (phenotypes). Ka ho hlahloba matšoao a liphatsa tsa lefutso ho pholletsa le genome eohle ho batho ba bangata, GWAS e hlalosa likamano tsa genotype-phenotype ka litlhahlobo tsa lipalo-palo tsa baahi. Mokhoa ona o fumana lits'ebetso tse pharaletseng ho batlisiseng mafu a batho le ho batlisisa liphatsa tsa lefutso tse sebetsang tse amanang le litšobotsi tse rarahaneng liphoofolong kapa limela.
Ho BMKGENE, re fana ka litsela tse peli tsa ho etsa GWAS ho baahi ba bangata: ho sebelisa Whole-Genome Sequencing (WGS) kapa ho khetha mokhoa o fokolitsoeng oa ho latellana ha liphatsa tsa lefutso, e leng Specific-Locus Amplified Fragment (SLAF) e entsoeng ka tlung. Leha WGS e lumellana le liphatsa tsa lefutso tse nyane, SLAF e hlaha e le mokhoa o mong o theko e tlaase bakeng sa ho ithuta baahi ba bangata ba nang le liphatsa tsa lefutso tse telele, e fokotsa litšenyehelo tsa ho latellana ka katleho, ha e ntse e netefatsa katleho e phahameng ea ho sibolla letšoao la liphatsa tsa lefutso.
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Tatellano ea Genome ea Limela/Liphoofolo ka Botlalo
Mokhoa oa ho Latela Li-Genome ka Botlalo (WGS) ke mokhoa o sebelisoang ho fumana tatellano ea DNA ea genome ea ntho e phelang ka nako e le 'ngoe.
Hangata, tšebeletso e arotsoe ka lihlopha tse peli tse fapaneng ho latela boteng ba genome ea referense:
- De novotatellano eohle ea liphatsa tsa lefutso.Boemong bona, genome e lokelang ho hlophisoa ha e na genome ea litšupiso e fumanehang, 'me ka lebaka leo, sepheo sa tatellano ena ke ho e hlahisa (kapa ho ntlafatsa e seng e ntse e le teng). Mokhoa ona o hloka ho sebelisa bobeli, data ea Illumina le tatellano ea nako e telele ho ntlafatsa kopano ea genome ka ho theha ho kopana pakeng tsa ho bala.
- Ho hlophisa bocha.E bua ka tatellano eohle ea liphatsa tsa lefutso tsa mefuta e fapaneng ea liphoofolo tse nang le liphatsa tsa lefutso tse tsejoang. Motheong ona, liphapang tsa liphatsa tsa lefutso tsa batho ka bomong kapa baahi li ka tsejoa haholoanyane.
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Liphatsa tsa lefutso tsa ho iphetola ha lintho
Thuto ea Liphatsa tsa Lefutso ke tšebeletso e felletseng ea tatellano e etselitsoeng ho fana ka tlhaloso e nang le temohisiso ea phetoho ka har'a sehlopha se seholo sa batho ka bomong, ho latela mefuta e fapaneng ea liphatsa tsa lefutso, ho kenyeletsoa le li-SNP, InDels, SV, le CNV. Tšebeletso ena e kenyelletsa litlhahlobo tsohle tsa bohlokoa tse hlokahalang ho hlakisa liphetoho tsa phetoho le litšobotsi tsa liphatsa tsa lefutso tsa baahi, ho kenyeletsoa le litekolo tsa sebopeho sa baahi, mefuta e fapaneng ea liphatsa tsa lefutso le likamano tsa phylogenetic. Ho feta moo, e kenella lithutong tse mabapi le phallo ea liphatsa tsa lefutso, e nolofalletsang likhakanyo tsa boholo bo sebetsang ba baahi le nako ea ho fapana. Lithuto tsa thuto ea liphatsa tsa lefutso tsa phetoho li fana ka temohisiso ea bohlokoa mabapi le tšimoloho le ho ikamahanya ha mefuta.
Ho BMKGENE, re fana ka litsela tse peli tsa ho etsa lithuto tsa liphatsa tsa lefutso tsa ho iphetola ha lintho ho baahi ba bangata: ho sebelisa tatellano ea liphatsa tsa lefutso tse felletseng (WGS) kapa ho khetha mokhoa oa tatellano ea liphatsa tsa lefutso tse fokolitsoeng, e leng Specific-Locus Amplified Fragment (SLAF) e ntlafalitsoeng ka tlung. Leha WGS e lumellana le liphatsa tsa lefutso tse nyane, SLAF e hlaha e le mokhoa o mong o theko e tlaase bakeng sa ho ithuta baahi ba bangata ba nang le liphatsa tsa lefutso tse telele, e leng se fokotsang litšenyehelo tsa tatellano ka katleho.
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Genomics e Bapisang
Papiso ea genomics e kenyelletsa tlhahlobo le papiso ea tatellano eohle ea genome le meaho har'a mefuta e fapaneng. Tšimo ena e batla ho senola phetoho ea mefuta, ho hlalosa mesebetsi ea liphatsa tsa lefutso, le ho hlakisa mekhoa ea taolo ea liphatsa tsa lefutso ka ho khetholla meaho le likarolo tsa tatellano tse bolokiloeng kapa tse fapaneng ho pholletsa le lintho tse phelang tse fapaneng. Thuto e felletseng ea papiso ea genomics e kenyelletsa litlhahlobo tse kang malapa a liphatsa tsa lefutso, nts'etsopele ea ho iphetola ha lintho, liketsahalo tsa ho pheta-pheta ha genome eohle, le tšusumetso ea likhatello tse khethiloeng.
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Kopano ea Genome e Thehiloeng ho Hi-C
Hi-C ke mokhoa o reretsweng ho hapa sebopeho sa chromosome ka ho kopanya dikamano tse itshetlehileng hodima bohaufi le tatellano e phahameng ya ho fetiswa ha di-chromosome. Matla a dikamano tsena ho dumelwa hore a amana hampe le sebaka sa mmele ho di-chromosome. Ka hona, data ya Hi-C e sebedisetswa ho tataisa ho bokellana, ho hlophisa le ho shebana ha di-tatelano tse bokelletsweng ho genome e hlophisitsweng le ho di tiisa hodima palo e itseng ya di-chromosome. Theknoloji ena e matlafatsa kopano ya genome ya boemo ba chromosome ha ho se na 'mapa wa lefutso o thehilweng hodima baahi. Genome e nngwe le e nngwe e hloka Hi-C.
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Semela / Phoofolo De Novo Genome Sequencing
De NovoHo latellana ho bolela kaho ea genome eohle ea mofuta ho sebelisoa mahlale a ho latellana ha ho se na genome ea litšupiso. Kenyelletso le kamohelo e pharaletseng ea tatellano ea moloko oa boraro, e nang le ho bala hole, li ntlafalitse kopano ea genome haholo ka ho eketsa ho kopana lipakeng tsa ho bala. Ntlafatso ena e bohlokoa haholo ha ho sebetsanoa le li-genome tse phephetsang, tse kang tse bontšang heterozygosity e phahameng, karolelano e phahameng ea libaka tse pheta-phetoang, li-polyploid, le libaka tse nang le likarolo tse pheta-phetoang, litaba tse sa tloaelehang tsa GC, kapa ho rarahana ho hoholo hoo hangata ho sa kopaneng hantle ho sebelisoa tatellano ea ho bala ho khutšoanyane feela.
Tharollo ea rona ea sebaka se le seng e fana ka lits'ebeletso tse kopaneng tsa tatellano le tlhahlobo ea bioinformatic e fanang ka genome ea boleng bo holimo e kopantsoeng. Phuputso ea pele ea genome ka Illumina e fana ka likhakanyo tsa boholo ba genome le ho rarahana ha eona, 'me tlhahisoleseling ena e sebelisoa ho tataisa mohato o latelang oa tatellano e baloang nako e telele ka PacBio HiFi, e lateloa kede novokopano ea li-contig. Tšebeliso e latelang ea kopano ea HiC e nolofalletsa ho khomarela li-contig ho genome, ho fumana kopano ea boemo ba chromosome. Qetellong, genome e hlalosoa ka ho bolela esale pele liphatsa tsa lefutso le ka ho latellana ha liphatsa tsa lefutso tse hlahisitsoeng, ho sebelisoa li-transcriptome tse nang le lipalo tse khutšoanyane le tse telele.
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Tatellano ea Exome ea Motho ka Botlalo
Ho latellana ha exome ea Motho ka Moka (hWES) ho ananeloa haholo e le mokhoa o theko e tlaase le o matla oa ho latellana bakeng sa ho khetholla liphetoho tse bakang mafu. Leha e le karolo ea 1.7% feela ea genome eohle, li-exons li bapala karolo ea bohlokoa ka ho bonahatsa ka kotloloho boemo ba mesebetsi eohle ea protheine. Haholo-holo, genome ea motho, ho feta 85% ea liphetoho tse amanang le mafu tse bonahalang libakeng tsa ho ngola liprotheine. BMKGENE e fana ka tšebeletso e felletseng le e feto-fetohang ea ho latellana ha exome ea motho ka maano a mabeli a fapaneng a ho hapa exon a fumanehang ho fihlela lipheo tse fapaneng tsa lipatlisiso.
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Tatellano ea Likaroloana tse Entsoeng ka Sebaka se Ikhethang (SLAF-Seq)
Mokhoa ona o ntshetswang pele ke BMKGene ka boikemelo, o ka arolwa ka hara tatellano ya genome e fokoditsweng. O ntlafatsa sete ya enzyme ya thibelo bakeng sa projeke e nngwe le e nngwe. Sena se netefatsa tlhahiso ya palo e kgolo ya di-tag tsa SLAF (dibaka tsa 400-500 bps tsa genome tse arolwang ka tatellano) tse arolwang ka ho lekana ho pholletsa le genome ha ka katleho di ntse di qoba dibaka tse pheta-phetoang, ka hona di netefatsa ho sibollwa ha matshwao a lefutso a molemo ka ho fetisisa.
E fana ka tlhahlobo e potlakileng ea liphatsa tsa lefutso 'me e beha motheo oa ho sibolloa ha liphatsa tsa lefutso tse sebetsang kapa tlhahlobo ea ho iphetola ha lintho e fokotsang litšenyehelo ka sampole ha e ntse e boloka bokhoni ba ho sibolloa ha matšoao a liphatsa tsa lefutso. RRGS e finyella sena ka ho sila DNA ka li-enzyme tse thibelang le ho tsepamisa maikutlo ho mefuta e itseng ea boholo ba likaroloana, ka hona e hlophisa karoloana feela ea genome. Har'a mekhoa e fapaneng ea RRGS, Specific-Locus Amplified Fragment Sequencing (SLAF) ke mokhoa o ka fetoloang le oa boleng bo holimo.
