Iimveliso

I-Specific-Locus Amplified Fragment Sequencing (SLAF-Seq)

I-genotyping ephezulu, ngokukodwa kwinani elikhulu labantu, linyathelo elisisiseko kwizifundo zombutho wemfuza, obonelela ngesiseko sofuzo sofuzo olusebenzayo, uhlalutyo lwe-evolution, njl. Endaweni yolandelelwano olunzulu lwe-genome kwakhona, ukunciphisa ukumelwa kwe-genome sequencing (RRGS). ) yaziswa ukuze kuncitshiswe iindleko zolandelelwano ngesampulu nganye, ngeli lixa kugcinwe ukusebenza kakuhle ekufumaneni uphawu lwemfuzo.Oku kudla ngokuphunyezwa ngokutsala isiqwenga sothintelo ngaphakathi koluhlu lobungakanani obunikiweyo, olubizwa ngokuba yithala leencwadi lokumelwa elincitshisiweyo (RRL).I-Specific-locus amplified fragment sequencing (SLAF-Seq) sisicwangciso esiziphuhlisileyo se-SNP genotyping kunye okanye ngaphandle kwe-genome yesalathiso.
Iqonga: I-Illumina NovaSeq Platform

Qhagamshelana nathi

Iinkcukacha zeNkonzo

Iziphumo zeDemo

Iimpapasho Ezikhoyo

Iinkcukacha zeNkonzo

Inkqubo yobuGcisa

Ukuhamba komsebenzi

Iinkonzo eziluncedo

Ukufunyanwa kwamanqaku aphezulu-Itekhnoloji yokulandelelanisa itekhnoloji ephezulu inceda iSLAF-Seq ekufumaneni amakhulu amawaka ethegi ngaphakathi kwegenome yonke.

Ukuxhomekeka okuphantsi kwi-genome- Ingasetyenziswa kwiintlobo ngeentlobo okanye ngaphandle kwe-genome yesalathiso.

Uyilo lwesikimu esiguqukayo- I-enzyme enye, i-enzyme emibini, i-digestion ye-enzyme eninzi kunye neentlobo ezahlukeneyo ze-enzymes, zonke zinokukhethwa ukuhlangabezana neenjongo ezahlukeneyo zophando okanye iintlobo.Uvavanyo lwangaphambili kwi-silico lusetyenziselwa ukuqinisekisa uyilo olululo lwe-enzyme.

Ukwetyisa okusebenzayo kwe-enzymatic- Uvavanyo lwangaphambili lwenziwa ukuze kulungiswe iimeko, okwenza uvavanyo olusemthethweni luzinze kwaye luthembekile.Ubuchule bokuqokelelwa kwamaqhekeza kungafikelela ngaphezulu kwama-95%.

Iithegi zeSLAF ezisasazwe ngokulinganayo- Iithegi ze-SLAF zisasazwa ngokulinganayo kuzo zonke iichromosome ukuya kowona mlinganiselo mkhulu, ukuzuza umndilili we-1 SLAF nge-4 kb.

Ukuphepha okusebenzayo kokuphindaphinda- Ulandelelwano oluphindaphindiweyo kwidatha ye-SLAF-Seq iyancipha ukuya ngaphantsi kwe-5%, ngakumbi kwiintlobo ezinezinga eliphezulu lokuphindaphinda, njengengqolowa, umbona, njl.

Amava abanzi-Ngaphezu kwama-2000 eeprojekthi ezivaliweyo ze-SLAF-Seq kumakhulu eentlobo ezigubungela izityalo, izilwanyana ezincancisayo, iintaka, izinambuzane, izinto eziphilayo zasemanzini, njl.

Ukuziphuhlisa ngokwakho ukuhamba komsebenzi we-bioinformatiki-I-workflow edibeneyo ye-bioinformatic ye-SLAF-Seq yaphuhliswa yi-BMKGENE ukuqinisekisa ukuthembeka kunye nokuchaneka kwemveliso yokugqibela.

IiNgcaciso zeNkonzo

Iqonga	Conc.(ng/gl)	Iyonke (ug)	OD260/280
Illumina NovaSeq	>35	>1.6(Umqulu>15μl)	1.6-2.5

Qaphela: Iisampulu ezintathu, nganye inezikimu ze-enzyme ezintathu, ziya kwenzelwa uvavanyo lwangaphambili.

Isicwangciso esiCetyisiweyo solandelelwano

Ubunzulu bolandelelwano: 10X/Tag

Ubungakanani beGenome	Kucetyiswa iithegi zeSLAF
< 500 Mb	100K okanye WGS
500 Mb- 1 Gb	100 K
I-1 Gb -2 Gb	200 K
Iigenomes ezinkulu okanye ezintsonkothileyo	300 - 400K

Usetyenziso

Kucetyiswa

Umlinganiselo Wabemi

Isicwangciso solandelelwano nobunzulu

Ubunzulu

Inombolo yethegi

I-GWAS

Inombolo yesampuli ≥ 200

10X

Ngoku ka

ubungakanani begenome

I-Genetic Evolution

Umntu ngamnye

iqela elincinane ≥ 10;

iisampulu zizonke ≥ 30

10X

Unikezelo lweSampuli eCetyisiweyo

Isitya: 2 ml centrifuge tube

Kwiisampulu ezininzi, sincoma ukuba ungayigcini kwi-ethanol.

Iisampulu zokufakwa ileyibhile: Iisampulu kufuneka zibhalwe ngokucacileyo kwaye zifane nefom yesampulu yolwazi engenisiweyo.

Ukuthumela: Umkhenkce owomileyo: Iisampulu kufuneka zipakishwe kwiingxowa kuqala kwaye zingcwatywe kwindawo eyomileyo.

Ukuhanjiswa kwenkonzo

Isampulu ye-QC

Ulingo lokulinga

SLAF-uvavanyo

ULungiselelo lweThala leeNcwadi

Ukulandelelanisa

Uhlalutyo lwedatha

Iinkonzo zasemva kokuthengiswa

Ngaphambili: I-DNA / RNA Ukulandelelana - i-Nanopore Sequencer

Okulandelayo: Ulandelelwano lwe-Exome yoMntu

1. Izibalo zeziphumo zemephu

2. Uphuhliso lophawu lweSLAF

3. Ukwahluka kwenkcazo

Unyaka	Ijenali	IF	Isihloko	Usetyenziso
2022	Unxibelelwano lwendalo	17.694	Isiseko se-genomic se-giga-chromosomes kunye ne-giga-genome ye-peony yomthi I-Paeonia ostii	I-SLAF-GWAS
2015	Ingcali yePhytologist entsha	7.433	Imizila yeenyawo zasekhaya ixhasa imimandla ye-genomic ebalulekileyo kwi-agronomic iimbotyi zesoya	I-SLAF-GWAS
2022	Ijenali yoPhando oluPhezulu	12.822	Genome-wide-wide artificial introgressions of Gossypium barbadense into G. hirsutum bonisa i-loci ephezulu yokuphucula ngaxeshanye umgangatho we-cotton fiber kunye nesivuno iimpawu	I-SLAF-Evolutionary genetics
2019	Isityalo seemolekyuli	10.81	Uhlalutyo lweGenomic yabemi kunye neNdibano yaseDe Novo ityhila iMvelaphi yeWeedy Irayisi njengoMdlalo wokuZiphendukela kwemvelo	I-SLAF-Evolutionary genetics
2019	Imfuzo yendalo	31.616	Ukulandelelana kweGenome kunye neentlobo ezahlukeneyo zemfuza ye-carp eqhelekileyo, i-Cyprinus carpio	Imephu ye-SLAF-Linkage
2014	Imfuzo yendalo	25.455	I-genome ye-peanut elinywayo inika ukuqonda kwi-karyotypes ye-legume, i-polyploid indaleko kunye nokufuywa kwezityalo.	Imephu ye-SLAF-Linkage
2022	Ijenali yeBiyoteknoloji yeZityalo	9.803	Ukuchongwa kwe-ST1 kutyhila ukhetho olubandakanya ukuhamba ngeenyawo kwimophology yembewu kunye nomxholo we-oyile ngexesha lokufuywa kweembotyi zesoya	Uphuhliso lwe-SLAF-Marker
2022	Ijenali yaMazwe ngaMazwe yeNzululwazi yeMolekyuli	6.208	Ukuchongwa kunye noPhuhliso lweMakishi yeDNA yeWheat-Leymus mollis 2Ns (2D) Ukutshintshwa kweKhromosome yeDisomic	Uphuhliso lwe-SLAF-Marker