Specific-Locus Amplified Fragment Sequencing (SLAF-Seq)
Lintlha tsa Tšebeletso
Sekema sa Tekheniki
Phallo ea mosebetsi
Melemo ea Tšebeletso
Tšebeliso e ntle ea ho sibolla lesupa- Theknoloji ea tatellano ea maemo a holimo e thusa SLAF-Seq ho fumana li-tag tse likete tse makholo ka har'a genome eohle.
Ho itšetleha ho tlase ho genome- E ka sebelisoa ho mefuta e nang le genome kapa ntle le eona.
Moralo oa leano le feto-fetohang- Enzyme e le 'ngoe, li-enzyme tse peli, tšilo ea li-enzyme tse ngata le mefuta e fapaneng ea li-enzyme, kaofela li ka khethoa ho fana ka sepheo kapa mefuta e fapaneng ea lipatlisiso.Tlhahlobo ea pele ho silico e sebelisetsoa ho netefatsa moralo o nepahetseng oa enzyme.
Tšilo ea lijo e sebetsang hantle- Teko ea pele e ile ea etsoa ho ntlafatsa maemo, e leng se etsang hore teko e hlophisitsoeng e be e tsitsitseng le e ka tšeptjoang.Bokhoni ba ho bokella likaroloana bo ka fihlella ho feta 95%.
Li-tag tsa SLAF tse ajoang ka ho lekana- Li-tag tsa SLAF li ajoa ka ho lekana ho li-chromosome tsohle ho isa tekanyong e kholo, ho fihlela karolelano ea 1 SLAF ka 4 kb.
Ho qoba ho pheta-pheta ka katleho- Tatelano e pheta-phetoang ho data ea SLAF-Seq e fokotsehile ho ea tlase ho 5%, haholo mefuta e nang le maemo a holimo a pheta-phetoang, joalo ka koro, poone, joalo-joalo.
Phihlelo e pharaletseng-Merero e fetang 2000 e koetsoeng ea SLAF-Seq ho makholo a mefuta e koahelang limela, liphoofolo tse anyesang, linonyana, likokoanyana, libopuoa tsa metsing, jj.
Boikemisetso ba ts'ebetso ea bioinformatics e tsoetseng pele- Ts'ebetso e kopaneng ea bioinformatic workflow bakeng sa SLAF-Seq e ntlafalitsoe ke BMKGENE ho netefatsa ho ts'epahala le ho nepahala ha tlhahiso ea ho qetela.
Litlhaloso tsa Tšebeletso
| Sethala | Conc.(ng/gl) | Kakaretso (ug) | OD260/280 |
| Illumina NovaSeq | >35 | >1.6(Moqolo>15μl) | 1.6-2,5 |
Recommended Sequencing Strategy
Botebo ba tatellano: 10X/Tag
| Boholo ba Genome | Khothalletsoang SLAF Tags |
| <500 Mb | 100K kapa WGS |
| 500 Mb- 1 Gb | 100 K |
| 1 Gb -2 Gb | 200 K |
| Liphatsa tsa lefutso tse kholo kapa tse rarahaneng | 300 - 400K |
| Lisebelisoa
| E khothalelitsoe Palo ea Baahi
| Leano la tatellano le botebo
| |
| Botebo
| Nomoro ea Tag
| ||
| GWASE
| Nomoro ea mohlala ≥200
| 10X
|
Ho latela boholo ba genome
|
| Phetoho ea Liphatsa tsa lefutso
| Batho ka bomong sehlopha se senyenyane ≥ 10; kakaretso ea lisampole ≥30
| 10X
| |
Tlhahiso ea Mehlala e khothalelitsoeng
Setshelo: 2 ml centrifuge tube
Bakeng sa lisampole tse ngata, re khothaletsa ho se boloke ethanol.
Mohlala oa ho ngola: Mehlala e hloka ho ngoloa ka ho hlaka le ho tšoana le foromo ea sampole e rometsoeng.
Thomello: Leqhoa le omileng: lisampole li hloka ho kenngoa ka mekotleng pele ebe li patoa ka leqhoa le omileng.
Ts'ebetso ea ts'ebetso ea litšebeletso
Mohlala oa QC
Teko ea sefofane
SLAF-teko
Tokiso ea Laebrari
Tatelano
Tlhahlobo ea Lintlha
Litšebeletso tsa ka mor'a thekiso
1. Lipalopalo tsa sephetho sa 'mapa
2. Ntlafatso ea matšoao a SLAF
3. Phapang tlhaloso
| Selemo | Journal | IF | Sehlooho | Lisebelisoa |
| 2022 | Lipuisano tsa tlhaho | 17.694 | Motheo oa genomic oa giga-chromosomes le giga-genome ea sefate sa peony Paeonia ostii | SLAF-GWAS |
| 2015 | Setsebi se Secha sa Phytology | 7.433 | Mehato ea lapeng e tiisa libaka tsa genomic tsa bohlokoa ba temo ho soya | SLAF-GWAS |
| 2022 | Journal of Advanced Research | 12.822 | Genome-wide artificial introgressions of Gossypium barbadense into G. hirsutum hlahisa loci e phahameng bakeng sa ntlafatso ea nako e le 'ngoe ea boleng ba fiber ea k'hothone le chai makhabane | SLAF-Evolutionary liphatsa tsa lefutso |
| 2019 | Semela sa Molek'hule | 10.81 | Population Genomic Analysis le Kopano ea De Novo e Senola Tšimoloho ea Weedy Raese e le Papali ea Tsoelo-pele | SLAF-Evolutionary liphatsa tsa lefutso |
| 2019 | Nature Genetics | 31.616 | Tatelano ea liphatsa tsa lefutso le mefuta-futa ea liphatsa tsa lefutso tsa carp e tloaelehileng, Cyprinus carpio | 'Mapa oa SLAF-Linkage |
| 2014 | Nature Genetics | 25.455 | Genome ea peanut e lengoang e fana ka temohisiso ka li-karyotypes tsa legume, polyploid. ho iphetola ha lintho le ho rua lijalo. | 'Mapa oa SLAF-Linkage |
| 2022 | Plant Biotechnology Journal | 9.803 | Ho tsebahatsa ST1 ho senola khetho e kenyelletsang ho palama ha morphology ea peo le oli ea mafura nakong ea tlhahiso ea soya | nts'etsopele ea SLAF-Marker |
| 2022 | International Journal of Molecular Sciences | 6.208 | Boitsebiso le Ntlafatso ea Letšoao la DNA bakeng sa Wheat-Leymus mollis 2Ns (2D) Phapang ea Disomic Chromosome | nts'etsopele ea SLAF-Marker |
