Kev ntsuam xyuas Segregant
Kev Pabcuam Zoo
Takagi li al., The plant journal, 2013
● Qhov tseeb hauv cheeb tsam: Sib tov ntau nrog 30+30 txog 200+200 tus neeg kom txo qis suab nrov tom qab;non-synonymous mutatants-raws li tus neeg sib tw cheeb tsam twv ua ntej.
● Kev tshuaj xyuas dav dav: Cov neeg sib tw sib sib zog nqus gene muaj nuj nqi annotation, suav nrog NR, SwissProt, GO, KEGG, COG, KOG, thiab lwm yam.
● Lub sijhawm hloov pauv sai dua: Kev hloov pauv cov noob nrawm hauv 45 hnub ua haujlwm.
● Kev paub dav dav: BMK tau pab txhawb ntau txhiab tus yam ntxwv hauv zos, suav nrog ntau hom qoob loo, cov khoom hauv dej, hav zoov, paj, txiv hmab txiv ntoo, thiab lwm yam.
Service Specifications
Cov pejxeem:
Segregating progeny ntawm niam txiv nrog opposing phenotypes.
Piv txwv li F2 progeny, Backcrossing (BC), Recombinant inbred kab (RIL)
Sib tov pas
Rau cov yam ntxwv zoo: 30 txog 50 tus neeg (tsawg kawg 20) / ntau
Rau kev ua kom muaj nuj nqis: sab saum toj 5% mus rau 10% cov tib neeg uas muaj phenotypes nyob rau hauv tag nrho cov pejxeem (tsawg kawg 30 + 30).
Pom zoo sequencing qhov tob
Tsawg kawg yog 20X / niam txiv thiab 1X / tus xeeb ntxwv ib tus neeg (xws li rau cov xeeb ntxwv sib xyaw pas dej ua ke ntawm 30 + 30 tus neeg, qhov sib txuas qhov tob yuav yog 30X ib qho loj)
Bioinformatics tsom xam
● Tag nrho genome resequencing
● Kev ua cov ntaub ntawv
● SNP/Indel hu
● Kev tshuaj ntsuam xyuas cheeb tsam
● Cov neeg sib tw gene muaj nuj nqi annotation
Sample Requirements thiab Delivery
Cov Qauv Xav Tau:
Nucleotides:
| gDNA qauv | Cov ntaub so ntswg |
| Concentration: ≥30 ng / μl | Loj: 1-2 g |
| Ntim: ≥2 μg (Volumn ≥15 μl) | Tsiaj txhu: 0.5-1 g |
| Purity: OD260/280 = 1.6-2.5 | Cov ntshav tag nrho: 1.5 ml |
Kev Pabcuam Ua Haujlwm Flow
Kev sim tsim
Qauv xa tuaj
RNA rho tawm
Kev tsim kho tsev qiv ntawv
Sequencing
Cov ntaub ntawv tsom xam
Kev pab cuam tom qab muag
1.Association analysis base on Euclidean Distance (ED) los txheeb xyuas cov neeg sib tw hauv cheeb tsam.Hauv daim duab hauv qab no
X-axis: Chromosome tooj;Txhua lub dot sawv cev rau tus nqi ED ntawm SNP.Cov kab Dub sib haum rau haum ED tus nqi.Tus nqi ED ntau dua qhia tias muaj kev sib koom ua ke tseem ceeb ntawm qhov chaw thiab phenotype.Red dash kab sawv cev rau qhov pib ntawm kev koom tes tseem ceeb.
2.Association tsom xam raws li tsis muaj SNP-index
X-axis: Chromosome tooj;Txhua dot sawv cev rau SNP-index tus nqi.Cov kab dub sawv rau haum SNP-index tus nqi.Qhov loj dua tus nqi yog, qhov tseem ceeb ntawm lub koom haum yog.
BMK Cas
Qhov loj-txhim kho qhov zoo ntawm qhov chaw Fnl7.1 encodes lig embryogenesis muaj protein ntau txuam nrog txiv hmab txiv ntoo caj dab ntev hauv dib.
Luam tawm: Plant Biotechnology Journal, xyoo 2020
Sequence Strategy:
Cov niam txiv (Jin5-508, YN): Tag nrho cov genome resequencing rau 34 × thiab 20 ×.
DNA pas dej (50 Ntev-caj dab thiab 50 luv-caj dab): Rov ua dua rau 61 × thiab 52 ×
Cov txiaj ntsig tseem ceeb
Hauv txoj kev tshawb no, cais cov pejxeem (F2 thiab F2: 3) tau tsim los ntawm kev hla lub caj dab ntev ntev Jin5-508 thiab caj dab luv YN.Ob lub pas dej DNA tau tsim los ntawm 50 tus neeg lub caj dab ntev thiab 50 tus neeg caj dab luv heev.Cov txiaj ntsig tseem ceeb QTL tau txheeb xyuas ntawm Chr07 los ntawm BSA tsom xam thiab cov qauv QTL daim ntawv qhia.Cov cheeb tsam tus neeg sib tw tau txo qis ntxiv los ntawm kev nplua-mapping, gene qhia kom muaj nuj nqis thiab kev sim hloov pauv, uas qhia cov noob tseem ceeb hauv kev tswj lub caj dab ntev, CsFnl7.1.Tsis tas li ntawd, polymorphism hauv CsFnl7.1 txhawb nqa cheeb tsam tau pom tias muaj feem cuam tshuam nrog cov lus qhia sib xws.Ntxiv phylogenetic tsom xam pom tias Fnl7.1 locus feem ntau yuav los ntawm Is Nrias teb.
 QTL-mapping nyob rau hauv BSA tsom xam los txheeb xyuas cov neeg sib tw cheeb tsam cuam tshuam nrog dib caj dab ntev |  LOD profiles ntawm dib caj dab ntev QTL txheeb xyuas ntawm Chr07 |
Xu, X., et al."Qhov loj-txhim kho qhov zoo ntawm qhov chaw Fnl7.1 encodes lig embryogenesis muaj protein ntau cuam tshuam nrog txiv hmab txiv ntoo caj dab ntev hauv dib."Plant Biotechnology Journal 18.7 (2020).
