Kev Txheeb Xyuas Bisulfite Sequencing (RRBS) Uas Txo Qhov Sawv Cev
Kev txo qis kev sawv cev bisulfite sequencing (RRBS) vam khom kev nplua nuj ntawm CpG kob-nplua nuj thaj chaw los ntawm MspI cleavage ua raws li kev xaiv qhov loj ntawm 200-500/600 bps fragments. Yog li ntawd, tsuas yog thaj chaw ze rau CpG kob raug sequenced, thaum cov uas muaj CpG kob deb raug tshem tawm ntawm kev tshuaj xyuas. Cov txheej txheem no, ua ke nrog bisulfite sequencing, tso cai rau kev kuaj pom siab ntawm DNA methylation, thiab txoj hauv kev sequencing, PE150, tsom mus rau qhov kawg ntawm cov ntxig es tsis yog nruab nrab, ua rau muaj kev ua haujlwm ntawm methylation profiling.
Cov txheej txheem no tau tshwm sim los ua lwm txoj hauv kev pheej yig thiab ua tau zoo rau Whole Genome Bisulfite Sequencing (WGBS) hauv kev tshawb fawb DNA methylation. Thaum WGBS muab kev nkag siab dav dav los ntawm kev tshuaj xyuas tag nrho cov genome ntawm qhov kev daws teeb meem ib qho, nws tus nqi siab tuaj yeem yog qhov txwv, uas RRBS ua kom txo qhov teeb meem no los ntawm kev xaiv kev tshuaj xyuas ib feem ntawm cov genome. Txoj kev no yog ib qho cuab yeej muaj nqis uas ua rau muaj kev tshawb fawb DNA methylation pheej yig thiab txhim kho kev paub txog cov txheej txheem epigenetic.
Cov yam ntxwv ntawm kev pabcuam
● Kev txheeb xyuas kab ke ntawm Illumina NovaSeq.
● Yuav tsum muaj ib qho genome siv ua piv txwv.
● Lambda DNA siv los saib xyuas qhov ua tau zoo ntawm kev hloov pauv bisulfite.
● Kev zom zaub mov zoo ntawm MspI kuj raug saib xyuas.
● Ob chav enzyme zom zaub mov rau cov qauv cog.
Cov Kev Pabcuam Zoo
●Tus nqi tsim nyog thiab ua tau zoo dua lwm txoj hauv kev rau WGBSNws ua rau kev tshuaj xyuas tuaj yeem ua tiav ntawm tus nqi qis dua thiab nrog cov qauv qis dua.
●Platform tiav:Peb muab kev pabcuam zoo heev ib zaug xwb los ntawm kev ua cov qauv, kev tsim tsev qiv ntawv, thiab kev txheeb xyuas cov ntaub ntawv mus rau kev tshuaj xyuas bioinformatics.
●Kev Paub Ntau YamPeb muaj kev vam meej thoob plaws ntau hom tsiaj sib txawv. BMKGENE coj ntau tshaj kaum xyoo ntawm kev paub dhau los, pab pawg neeg tshuaj xyuas kev txawj ntse, cov ntsiab lus tiav, thiab kev txhawb nqa tom qab muag zoo heev.
Cov Lus Qhia Txog Kev Pabcuam
| Tsev qiv ntawv | Txoj Kev Npaj Txog Sequencing | Cov ntaub ntawv tso tawm pom zoo | Kev tswj xyuas zoo |
| MspI zom thiab Bisulfite kho tsev qiv ntawv | Illumina PE150 | 8 Gb | Q30 ≥ 85% Kev hloov pauv ntawm Bisulfite > 99% Kev ua haujlwm txiav MspI > 95% |
Cov Qauv Yuav Tsum Tau Ua
| Kev Ntsuas (ng/µL) | Tag nrho cov nqi (µg) |
| |
| DNA genomic | ≥ 30 | ≥ 1 | Kev puas tsuaj lossis kev ua qias tuaj tsawg |
Kev Ua Haujlwm Pabcuam
Kev xa khoom qauv
Kev tsim kho tsev qiv ntawv
Kev txheeb xyuas kab ke
Kev tshuaj xyuas cov ntaub ntawv
Cov kev pab cuam tom qab muag
Kev tshuaj xyuas BI muaj xws li:
● Kev tswj xyuas qhov zoo ntawm cov ntaub ntawv raw;
● Kev kos duab rau cov genome siv;
● Kev nrhiav pom 5mC methylated bases thiab kev txheeb xyuas motif;
● Kev tshuaj xyuas ntawm kev faib tawm methylation thiab kev sib piv cov qauv;
● Kev Tshawb Fawb Txog Cov Cheeb Tsam Methylated Sib Txawv (DMRs);
● Kev piav qhia txog kev ua haujlwm ntawm cov noob caj noob ces uas cuam tshuam nrog DMRs.
Kev tswj xyuas zoo: kev zom zaub mov zoo (hauv kev kos duab genome)
Kev tswj xyuas zoo: kev hloov pauv bisulfite (hauv kev rho tawm cov ntaub ntawv methylation)
Daim ntawv qhia Methylation: 5mC methylation genome-thoob plaws kev faib tawm
Kev sib piv piv txwv: Kev tshuaj xyuas cov khoom tseem ceeb
Kev tshuaj xyuas thaj chaw sib txawv ntawm cov cheeb tsam Methylated (DMRs): daim ntawv qhia kub
Tshawb nrhiav kev nce qib ntawm kev tshawb fawb uas tau pab txhawb los ntawm BMKGene cov kev pabcuam sequencing genome bisulfite tag nrho los ntawm kev sau cov ntawv tshaj tawm.
Li, Z. et al. (2022) 'Kev tsim kho dua tshiab zoo rau hauv Leydig-zoo li hlwb los ntawm CRISPR activation thiab paracrine factors',PNAS Nexus, 1(4). doi: 10.1093/PNASNEXUS/PGAC179.
Tian, H. et al. (2023) 'Kev tshuaj xyuas DNA methylation thoob plaws genome ntawm lub cev muaj pes tsawg leeg hauv Suav teb monozygotic ntxaib',European Journal of Clinical Investigation, 53(11), p. ib 14055. doi: 10.1111/ECI.14055.
Wu, Y. et al. (2022) 'DNA methylation thiab qhov sib piv ntawm lub duav thiab lub duav: kev tshawb fawb txog kev sib raug zoo thoob plaws epigenome hauv cov menyuam ntxaib monozygotic Suav',Phau Ntawv Xov Xwm Txog Kev Tshawb Fawb Endocrinological, 45(12), pp. 2365–2376. doi: 10.1007/S40618-022-01878-4 ib.
