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Lihlahisoa

Representation Bisulfite Sequencing (RRBS)

Setšoantšo sa 84

Reduced Representation Bisulfite Sequencing (RRBS) e hlahile e le mokhoa o sa sebetseng hantle le o sebetsang hantle oa Whole Genome Bisulfite Sequencing (WGBS) lipatlisisong tsa DNA methylation. Le ha WGBS e fana ka lintlha tse felletseng ka ho hlahloba genome eohle ka qeto e le 'ngoe ea motheo, litšenyehelo tsa eona tse phahameng e ka ba ntho e fokotsang. RRBS ka maano e fokotsa phephetso ena ka ho khetha karolo e emelang ea genome. Mokhoa ona o ipapisitse le ntlafatso ea libaka tse ruileng tsa lihlekehleke tsa CpG ke MspI cleavage e lateloang ke khetho ea boholo ba likhechana tsa 200-500/600 bps. Ka lebaka leo, ke libaka tse haufi le lihlekehleke tsa CpG feela tse latellanang, athe tse nang le lihlekehleke tse hole tsa CpG li sa kengoa tlhahlobong. Ts'ebetso ena, e kopantsoeng le tatellano ea bisulfite, e lumella ho lemoha ho phahameng ka ho fetisisa ha DNA methylation, le mokhoa oa ho latellana, PE150, o lebisa tlhokomelo ka ho khetheha lipheletsong tsa ho kenya ho e-na le bohareng, ho eketsa katleho ea profiling ea methylation. RRBS ke sesebelisoa sa bohlokoahali se nolofalletsang lipatlisiso tsa DNA methylation tse theko e tlaase le ho ntlafatsa tsebo ea mekhoa ea epigenetic.


Lintlha tsa Tšebeletso

Bioinformatics

Sephetho sa Demo

Lingoliloeng tse hlahang

Likarolo tsa Tšebeletso

● E hloka genome ea referense.

● Lambda DNA e sebelisetsoa ho beha leihlo katleho ea phetoho ea bisulfite.

● MspI tshilo ya dijo e boetse e behilwe leihlo.

● Ho sila ha li-enzyme tse habeli bakeng sa lisampole tsa semela.

● Sequencing ho Illumina NovaSeq.

Melemo ea Tšebeletso

Mokhoa o mong o sebetsang hantle le o sebetsang hantle oa WGBS: ho nolofalletsa hore tlhahlobo e etsoe ka theko e tlase le ka litlhoko tse tlase tsa sampole.

Sethala se Felletseng:fana ka ts'ebeletso e ntle ea sebaka se le seng ho tloha ts'ebetsong ea sampole, kaho ea laeborari, le tatellano ea tlhahlobo ea bioinformatics.

Tsebo e Pharalletseng: ka merero ea tatellano ea RRBS e phethiloeng ka katleho mefuteng e fapaneng ea mefuta-futa, BMKGENE e tlisa phihlelo ea lilemo tse fetang leshome, sehlopha sa bahlahlobisisi ba nang le tsebo e phahameng, litaba tse feletseng, le tšehetso e babatsehang ea ka mor'a thekiso.

Litlhaloso tsa Tšebeletso

Laebrari

Leano la Tatelano

Tlhahiso ea data e khothalelitsoeng

Taolo ea boleng

MspI e silehileng le laeborari ea phekolo ea Bisulfite

Illumina PE150

8 Gb

Q30 ≥ 85%

Phetoho ea Bisulfite > 99%

Katleho ea ho itšeha ea MspI> 95%

Litlhoko tsa Mohlala

 

Khokahano (ng/µL)

Kakaretso (µg)

 

Genomic DNA

≥30

≥1

Phokotso e fokolang kapa tshilafatso

Phallo ea Mosebetsi oa Ts'ebeletso

tlhahiso ea mohlala

Mohlala oa thomello

Tokiso ea Laebrari

Kaho ea laebrari

Tatelano

Tatelano

Tlhahlobo ea lintlha

Tlhahlobo ea lintlha

Ka mor'a ho rekisa Litšebeletso

Litšebeletso tsa ka mor'a thekiso


  • E fetileng:
  • E 'ngoe:

  • Setšoantšo sa 85

    E kenyelletsa tlhahlobo e latelang:

    ● Taolo ea boleng ba tatellano e tala;

    ● Ho etsa 'mapa oa litšupiso tsa genome;

    ● Ho fumanoa ha 5mC methylated bases le boitsebiso ba motif;

    ● Tlhahlobo ea kabo ea methylation le papiso ea sampole;

    ● Tlhahlobo ea Libaka tse sa tšoaneng tsa Methylated (DMRs);

    ● Tlhaloso e sebetsang ea liphatsa tsa lefutso tse amanang le li-DMR.

    Taolo ea boleng: ts'ebetso ea tšilo ea lijo (ka 'mapa oa genome)

     

    图片86

     

    Taolo ea boleng: phetolo ea bisulfite (ka ho ntšoa ha lintlha tsa methylation)

     

    图片87

     

    'Mapa oa Methylation: 5mC methylation genome-wide distribution

    图片88

     

    Papiso ea mohlala: Tlhahlobo ea Karolo e ka Sehloohong

     

    图片89

     

    Tlhahlobo ea Libaka tse fapaneng tsa Methylated (DMRs): 'mapa oa mocheso

     

    图片90

     

     

    Lekola tsoelo-pele ea lipatlisiso e tsamaisoang ke litšebeletso tsa tatellano ea liphatsa tsa lefutso tsa BMKGene ka pokello e hlophisitsoeng ea lingoliloeng.

    Li, Z. et al. (2022) 'Ho tsosolosa botšepehi bo phahameng ka har'a lisele tse kang Leydig ka ts'ebetso ea CRISPR le lintlha tsa paracrine',PNAS Nexus, 1(4). doi: 10.1093/PNASNEXUS/PGAC179.

    Tian, ​​H. et al. (2023) 'Genome-wide DNA methylation analysis of body composition in Chinese monozygotic twins',European Journal of Clinical Investigation, 53(11), leq. e14055. doi: 10.1111/ECI.14055.

    Wu, Y. et al. (2022) 'DNA methylation le karo-karolelano ea thekeng-to-hip: thuto ea epigenome-wide association mafahla a monozygotic a Chaena',Journal of Endocrinological Investigation, 45(12), leqepheng la 2365-2376. doi: 10.1007/S40618-022-01878-4.

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