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Lihlahisoa

Specific-Locus Amplified Fragment Sequencing (SLAF-Seq)

High-throughput genotyping, haholo-holo ho baahi ba bangata, ke mohato oa bohlokoa lithutong tsa liphatsa tsa lefutso, tse fanang ka motheo oa liphatsa tsa lefutso bakeng sa ho sibolloa ha liphatsa tsa lefutso, tlhahlobo ea ho iphetola ha lintho, joalo-joalo Ho e-na le tatellano e tebileng ea liphatsa tsa lefutso, ho fokotseha ha tatellano ea liphatsa tsa lefutso (RRGS). ) e hlahisoa ho fokotsa litšenyehelo tsa tatellano ea sampuli ka 'ngoe, ha ho ntse ho boloka ts'ebetso e utloahalang ho sibolloeng ha lets'oao la lefutso.Hangata sena se finyelloa ka ho ntša sekhechana sa lithibelo ka har'a boholo bo fanoeng, se bitsoang laeborari ea boemeli bo fokolitsoeng (RRL).Specific-locus amplified fragment sequencing (SLAF-Seq) ke leano le iketselitseng la SNP genotyping ka kapa ntle le genome ea referense.
Sethala: Illumina NovaSeq Platform


Lintlha tsa Tšebeletso

Demo Results

Likhatiso Tse Tsoang

Lintlha tsa Tšebeletso

Sekema sa Tekheniki

111

Phallo ea mosebetsi

流程图

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

Ela hloko: Mehlala e meraro, e 'ngoe le e 'ngoe e na le merero e meraro ea li-enzyme, e tla etsoa bakeng sa teko ea pele.

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
Teko ea SLAF
Tokiso ea Laebrari
Tatelano
Tlhahlobo ea lintlha
Ka mor'a ho rekisa Litšebeletso

Mohlala oa QC

Teko ea sefofane

SLAF-teko

Tokiso ea Laebrari

Tatelano

Tlhahlobo ea Lintlha

Litšebeletso tsa ka mor'a thekiso


  • E fetileng:
  • E 'ngoe:

  • 1. Lipalopalo tsa sephetho sa 'mapa

    setšoantšo1

    A1

    2. Ntlafatso ea matšoao a SLAF

    A2

    3. Phapang tlhaloso

    A3

    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

    fumana qotso

    Ngola molaetsa wa hao mona mme o re romele wona

    Re romelle molaetsa oa hau: