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Taxanaha Jajabka La Weyneeyay ee Gaarka ah-Locus (SLAF-Seq)

Genotyping-sare-sare, gaar ahaan dadka tirada badan, waa tallaabo aasaasiga ah ee daraasadaha ururka hidde, kaas oo bixiya aasaaska hidde-helidda hiddo-wadaha functional, falanqaynta evolutionary, iwm. ) waxaa loo soo bandhigay si loo yareeyo qiimaha isku xigxiga ee muunad kasta, iyadoo la ilaalinayo waxtarka macquulka ah ee helitaanka calaamadeeyaha hidde-sideyaasha.Tan waxaa badanaa lagu gaaraa in la soo saaro jajab xaddidan oo ku dhex jira cabbirka cabbirka, kaas oo lagu magacaabo maktabad matalaad la dhimay (RRL).Taxanaha goos gooska gaarka ah ee goobta la kordhiyey (SLAF-Seq) waa istaraatiijiyad iskeed u horumarisay oo loogu talagalay SNP genotyping oo leh ama aan lahayn genome tixraaceed.
Platform: Illumina NovaSeq Platform


Faahfaahinta Adeegga

Natiijooyinka Demo

Daabacaada sifada

Faahfaahinta Adeegga

Qorshaha Farsamada

111

Socodka shaqada

流程图

Faa'iidooyinka Adeegga

Hufnaanta calaamadaynta sare- Tiknoolajiyada isku xigxiga ee sare-u-qaadka ah waxay ka caawisaa SLAF-Seq soo saarista boqollaal kun oo calaamado ah oo ku dhex jira genome-ga oo dhan.

Ku tiirsanaanta hoose ee genome- Waxaa lagu dabaqi karaa noocyada mid leh ama aan lahayn genome tixraac ah.

Naqshadaynta nidaamka dabacsan- Hal-enzyme, dual-enzyme, dheef-shiid kiimikaad badan iyo noocyo kala duwan oo enzymes ah, dhammaantood waa la dooran karaa si loo daboolo yoolalka cilmi-baarista ama noocyada kala duwan.Qiimaynta hore ee silico ayaa loo isticmaalaa si loo xaqiijiyo naqshadaynta enzyme ugu fiican.

Dheefshiidka enzymatic oo hufan- Tijaabo hore ayaa loo sameeyay si loo wanaajiyo xaaladaha, taas oo ka dhigaysa tijaabada rasmiga ah mid deggan oo la isku halayn karo.Hufnaanta ururinta jajabku waxay gaari kartaa in ka badan 95%.

Calaamadaha SLAF si siman loo qaybiyay- Calaamadaha SLAF waxay si siman ugu qaybsan yihiin dhammaan koromosoomyada illaa xadka ugu badan, iyagoo gaaraya celcelis ahaan 1 SLAF 4 kb.

Ka fogaanshaha waxtarka leh ee soo noqnoqdaIsku-xigta soo noqnoqda ee xogta SLAF-Seq ayaa hoos loo dhigay ilaa 5%, gaar ahaan noocyada leh heerka sare ee soo noqnoqda, sida sarreenka, galleyda, iwm.

Khibrad ballaadhan-In ka badan 2000 oo mashruuc SLAF-Seq ah oo xidhay boqolaal nooc oo daboolaya dhirta, naasleyda, shimbiraha, cayayaanka, noolaha-aqua-organisms, iwm.

Socod-shaqo bioinformatic ah oo iskeed u horumarisay- Socod-hawleed bioinformatic ah oo isku-dhafan oo loogu talagalay SLAF-Seq waxaa soo saaray BMKGENE si loo hubiyo isku halaynta iyo saxnaanta wax-soo-saarka kama-dambaysta ah.

 

Tilmaamaha Adeegga

 

Platform

Conc.(ng/gl)

Wadartaug)

OD260/280

Illumina NovaSeq

>35

>1.6(Qaybta>15μl)

1.6-2.5

Fiiro gaar ah: Saddex muunado, mid kastaa leh saddex nidaam oo ensaymes ah, ayaa loo samayn doonaa tijaabada hore.

Istaraatiijiyada isku xigxiga ee lagu taliyay

Qoto dheer ee isku xigxiga: 10X/Tag

Cabirka Genome

Tags SLAF lagu taliyay

< 500 Mb

100k ama WGS

500 Mb- 1 Gb

100 K

1 Gb -2 Gb

200 K

Giant ama genomes adag

300 - 400K

 

Codsiyada

 

lagu taliyay

Qiyaasta Dadweynaha

 

Istaraatiijiyada isku xigxiga iyo qoto dheer

 

Qoto dheer

 

Lambarka Tag

 

GWAS

 

Tusaalaha nambarka ≥ 200

 

10X

 

 

 

 

 

Sida laga soo xigtay

cabbirka genome

 

Evolution Hidde

 

Shakhsiyaadka mid kasta

koox-hoosaad ≥ 10;

muunado guud ≥ 30

 

10X

 

Tusaalaha keenista lagu taliyay

Weelka: 2 ml tuubo centrifuge

Inta badan shaybaarada, waxaan kugula talineynaa inaan lagu keydin ethanol.

Calaamadaynta Tusaalaha: Muunaduhu waxay u baahan yihiin in si cad loo calaamadiyo oo la mid ah foomka macluumaadka muunada ee la gudbiyay.

Shixnadda: Barafka qallalan: Muunadodu waxay u baahan yihiin in marka hore bac lagu shubo oo lagu aaso baraf qallalan.

Socodka Shaqada Adeegga

Tusaalaha QC
Tijaabada tijaabada
Tijaabada SLAF
Diyaarinta Maktabadda
Isku xigxiga
Falanqaynta xogta
Adeegyada iibka kadib

Tusaalaha QC

Tijaabada tijaabada

SLAF-tijaabo

Diyaarinta Maktabadda

Isku xigxiga

Falanqaynta Xogta

Adeegyada iibka kadib


  • Hore:
  • Xiga:

  • 1. Tirakoobka natiijada khariidada

    sawirka1

    A1

    2. Horumarinta calaamadaha SLAF

    A2

    3. Faahfaahin kala duwan

    A3

    Sannadka

    Joornaal

    IF

    Ciwaanka

    Codsiyada

    2022

    Isgaarsiinta dabeecadda

    17.694

    Asalkii genomic ee giga-chromosomes iyo giga-genome of peony geed

    Paeonia ostii

    SAF-GWAS

    2015

    Cilmi-nafsiga cusub

    7.433

    Raad-raacyada gurigu waxay ku xidhaan gobollada genomic ee muhiimadda beeralayda gudaha

    digirta

    SAF-GWAS

    2022

    Joornaalka Cilmi-baarista Sare

    12.822

    Genom-ballaaran hordhaca macmalka ah ee Gossypium barbadense galay G. hirsutum

    Muujin heer sare ah oo loogu talagalay hagaajinta tayada fiber-ka suufka ah iyo wax-soo-saarka

    sifooyinka

    SLAF-horumarka hiddaha

    2019

    Warshada Molecular

    10.81

    Falanqaynta Genomic Dadweynaha iyo Golaha De Novo ayaa Muujiyay Asalka Weedy

    Bariiska sida Ciyaar Horumarineed

    SLAF-horumarka hiddaha

    2019

    Hiddaha Dabeecada

    31.616

    Isku xigxiga genome iyo kala duwanaanshaha hidde-sidaha ee carp-ka caadiga ah, Cyprinus carpio

    Khariidadda SLAF-Likage

    2014

    Hiddaha Dabeecada

    25.455

    Hiddo-sidaha lawska la beeray waxa uu siinaya aragtida legume karyotypes, polyploid

    kobcinta iyo dalagyada dalagyada.

    Khariidadda SLAF-Likage

    2022

    Wargeyska Biotechnology ee dhirta

    9.803

    Aqoonsiga ST1 wuxuu muujinayaa xulashada ku lug leh garaacista qaab-dhismeedka abuurka

    iyo waxa ku jira saliidda inta lagu jiro dhaqashada soybean

    Horumarinta SLF-Marker

    2022

    Joornaalka Caalamiga ah ee Cilmiga Molecular

    6.208

    Aqoonsiga iyo Horumarinta Calaamadaha DNA ee Qamadiga-Leymus mollis 2Ns (2D)

    Beddelka Kala-duwan ee Chromosome

    Horumarinta SLF-Marker

    qaado xigasho

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