Rage Matsayin Bisulfite Sequencing (RRBS)
Rage Wakilci Bisulfite Sequencing (RRBS) ya fito a matsayin mai inganci mai tsada da ingantaccen madadin Tsarin Tsarin Halittu na Halitta (WGBS) a cikin binciken DNA methylation. Yayin da WGBS ke ba da cikakkun bayanai ta hanyar nazarin dukkan kwayoyin halitta a ƙudurin tushe guda ɗaya, ƙimar sa mai girma na iya zama abin iyakancewa. RRBS da dabara yana rage wannan ƙalubalen ta hanyar zaɓin nazarin sashin wakilci na kwayoyin halitta. Wannan dabarar ta dogara ne akan haɓaka yankuna masu wadatar tsibiri na CpG ta hanyar tsagewar MspI wanda ke biye da girman zaɓi na 200-500/600 bps. Sakamakon haka, kawai yankuna masu kusanci zuwa tsibiran CpG ne aka jera su, yayin da waɗanda ke da tsibiran CpG masu nisa ba a cire su daga binciken. Wannan tsari, haɗe tare da bisulfite sequencing, yana ba da damar gano babban ƙuduri na DNA methylation, da tsarin tsarin, PE150, yana mai da hankali musamman a kan ƙarshen abubuwan da aka saka maimakon tsakiya, yana ƙara haɓakar haɓakar methylation. RRBS kayan aiki ne mai mahimmanci wanda ke ba da damar bincike na DNA methylation mai tsada mai tsada da haɓaka ilimin hanyoyin epigenetic.
Siffofin Sabis
● Yana buƙatar nau'in kwayar halitta.
Ana amfani da DNA na Lambda don saka idanu da ingancin jujjuyawar bisulfite.
● Ana kuma lura da ingancin narkewar MspI.
● Narkewar enzyme sau biyu don samfuran shuka.
● Sequencing akan Illumina NovaSeq.
Amfanin Sabis
●Madaidaicin farashi da ingantaccen Madadi zuwa WGBS: ba da damar bincike da za'ayi a cikin ƙananan farashi kuma tare da ƙananan buƙatun samfurin.
●Cikakken Dandali:samar da kyakkyawan sabis na tsayawa ɗaya daga sarrafa samfurin, ginin ɗakin karatu, da jeri zuwa nazarin bioinformatics.
●Ƙwararren Ƙwararru: tare da RRBS jerin ayyukan da aka samu nasarar kammala a cikin nau'i-nau'i daban-daban, BMKGENE ya kawo shekaru goma na kwarewa, ƙungiyar bincike mai mahimmanci, cikakkun abun ciki, da kuma kyakkyawan goyon bayan tallace-tallace.
Ƙayyadaddun Sabis
| Laburare | Dabarun Jeri | Fitowar bayanai da aka ba da shawarar | Kula da inganci |
| MspI ta narke da ɗakin karatu na Bisulfite | Farashin PE150 | 8 gb | Q30 ≥ 85% Canjin Bisulfite> 99% MspI yanke yadda ya dace> 95% |
Samfuran Bukatun
| Hankali (ng/µL) | Jimlar adadin (µg) |
| |
| Genomic DNA | ≥ 30 | ≥ 1 | Ƙayyadaddun lalacewa ko gurɓatawa |
Gudun Aikin Sabis
Samfurin bayarwa
Gina ɗakin karatu
Jeri
Binciken bayanai
Bayan-sayar da sabis
Ya haɗa da bincike mai zuwa:
● Raw sequencing ingancin kula;
● Yin taswira zuwa ga kwayoyin halitta;
● Gano 5mC methylated sansanonin da gano motif;
● Nazarin rarraba methylation da kwatanta samfurin;
● Binciken Yankunan Methylated Daban-daban (DMRs);
● Bayanin aiki na kwayoyin halitta masu alaƙa da DMRs.
Gudanar da inganci: ingancin narkewa (a cikin taswirar genome)
Gudanar da inganci: juyawa bisulfite (a cikin hakar bayanan methylation)
Taswirar Methylation: 5mC methylation genome-fadi rarraba
Misali kwatankwacin: Babban Fahimtar Fassarar
Binciken Yankunan Methylated Daban-daban (DMRs): taswirar zafi
Bincika ci gaban bincike ta hanyar BMKGene gabaɗayan hidimomin bisulfite na genome ta hanyar tarin wallafe-wallafe.
Li, Z. et al. (2022) 'Sake tsarin ingantaccen aminci cikin sel kamar Leydig ta hanyar kunna CRISPR da abubuwan paracrine',PNAS Nexus, 1 (4). doi: 10.1093/PNASNEXUS/PGAC179.
Tian, H. et al. (2023) 'Binciken DNA methylation na DNA mai fa'ida game da abun da ke ciki a cikin tagwayen monozygotic na kasar Sin',Jaridar Turai na Binciken Clinical, 53 (11), shafi. ku 14055. doi: 10.1111/ECI.14055.
Wu, Y. et al. (2022) 'DNA methylation da kugu-zuwa-hip rabo: nazarin ƙungiyar epigenome-fadi a cikin tagwayen monozygotic na kasar Sin',Jaridar Endocrinological Investigation, 45 (12), shafi na 2365-2376. doi: 10.1007/S40618-022-01878-4.
