Tsarin Rarraba Kayayyaki na Musamman (SLAF-Seq)
Tsarin aiki
Sabis ɗin yana da wasu ƙira da aka riga aka tsara a cikin silico don tabbatar da mafi kyawun zaɓin enzyme akan shirya ɗakin karatu.
Tsarin Fasaha
Fasallolin Sabis
● Jerin abubuwa akan NovaSeq tare da PE150.
● Shirye-shiryen ɗakin karatu tare da yin amfani da barcode sau biyu, wanda ke ba da damar tattara sama da samfura 1000.
● Ba tare da la'akari da kwayar halittar da aka ambata ba:
Tare da tsarin halittar da aka ambata: gano SNP da InDel
Ba tare da nassoshi ba: tarin samfurin da gano SNP
● A cikina cikin silikiAna tantance haɗakar enzyme masu ƙuntatawa da yawa na matakin ƙira kafin a fara amfani da su don nemo waɗanda ke samar da rarraba iri ɗaya na alamun SLAF tare da kwayar halittar.
● A lokacin gwaji kafin a fara, ana gwada haɗakar enzyme guda uku a cikin samfura 3 don samar da ɗakunan karatu guda 9 na SLAF, kuma ana amfani da wannan bayanin don zaɓar haɗin enzyme mafi kyau don aikin.
Fa'idodin Sabis
●Gano Babban Alamar Halitta: Mun haɗa tsarin lambar barcode mai ƙarfi wanda ke ba da damar yin jerin gwanon jama'a a lokaci guda, da kuma faɗaɗawa takamaiman wurare, wanda ke inganta inganci, yana tabbatar da cewa lambobin tag sun cika buƙatun daban-daban na tambayoyin bincike daban-daban.
● Ƙananan Dogara ga Halittar Halitta: Ana iya amfani da shi ga nau'in halittu tare da ko ba tare da kwayar halittar da aka ambata ba.
●Tsarin Tsarin Mai Sauƙi: Ana iya zaɓar nau'ikan enzymes guda ɗaya, nau'ikan enzymes biyu, nau'ikan enzymes daban-daban, da nau'ikan enzymes daban-daban don biyan buƙatun bincike ko nau'ikan.
● Ingantaccen Inganci a Narkewar Enzymatic: Gudanar da wania cikin silikiTsarin da aka riga aka tsara da kuma gwajin da aka riga aka yi yana tabbatar da ingantaccen tsari tare da rarraba alamun SLAF daidai gwargwado akan chromosome (alamar SLAF 1/4Kb) da kuma rage jerin maimaitawa (<5%).
●Ƙwarewa Mai Zurfi: Muna kawo tarin gogewa ga kowane aiki, tare da tarihin rufe ayyukan SLAF-Seq sama da 5000 akan ɗaruruwan nau'ikan halittu, gami da tsire-tsire, dabbobi masu shayarwa, tsuntsaye, kwari, da halittun ruwa.
● Tsarin Aiki na Bioinformatic wanda aka haɓaka da kansa: Mun ƙirƙiro tsarin aiki na bioinformatic wanda aka haɗa don SLAF-Seq don tabbatar da inganci da daidaito na sakamakon ƙarshe.
Bayanin Sabis
| Nau'in nazari | Girman yawan jama'a da aka ba da shawarar | Dabarun tsara tsari | |
| Zurfin jerin alamun | Lambar alama | ||
| Taswirorin Halitta | Iyaye 2 da 'ya'ya sama da 150 | Iyaye: 20x WGS Ragewa: 10x | Girman kwayar halitta: <400 Mb: Ana ba da shawarar WGS <1Gb: Alamun 100K 1-2Gb:: Alamun 200K >2Gb: Alamun 300K Matsakaicin alamun 500k |
| Nazarin Ƙungiyoyin Halittar Jini (GWAS) | ≥ samfura 200 | 10x | |
| Juyin Halittar Halitta | ≥samfura 30, tare da sama da samfura 10 daga kowace ƙungiya | 10x | |
Bukatun Sabis
Mayar da hankali ≥ 5 ng/µL
Jimlar adadin ≥ 80 ng
Gel na Agarose: babu ko iyakancewar lalacewa ko gurɓatawa
Isarwa Samfurin da Aka Ba da Shawara
Kwantena: bututun centrifuge 2 ml
(Ga yawancin samfuran, muna ba da shawarar kada a adana a cikin ethanol)
Lakabi da Samfura: Ya kamata a yi wa samfuran lakabi a sarari kuma su yi daidai da fom ɗin bayanin samfurin da aka gabatar.
Jigilar Kaya: Busasshen Kankara: Da farko ana buƙatar a saka samfuran a cikin jakunkuna sannan a binne su a cikin busasshen kankara.
Tsarin Aikin Sabis
Samfurin QC
Gwajin gwaji
gwajin SLAF
Shirye-shiryen Laburare
Jerin abubuwa
Binciken Bayanai
Ayyukan Bayan Sayarwa
Binciken bioinformatical ɗinmu ya ƙunshi:
QC na bayanai da kuma rage bayanai don cire karanta-riba na N, karanta adaftar ko karanta-ƙaramar inganci.
Kulawa ta biyu ta inganci don duba rarrabawar tushe, ingancin jerin abubuwa da kuma kimanta bayanai, amma kuma don duba ingancin narkewar abinci da kuma abubuwan da aka saka.
Da zarar an tabbatar da sakamakon, akwai zaɓuɓɓuka guda biyu:
- Taswirar zuwa ga genome mai ma'ana
- Ba tare da kwayar halittar da aka ambata ba: tarin kwayoyin halitta
Bayan haka, ana amfani da nazarin alamun SLAF don yin wasu nau'ikan kira don taimakawa tare da gano alamar: SNP, InDel, SNV, kiran CV da bayanin kula
Rarraba alamun SLAF akan ƙwayoyin halitta:
Rarraba SNPs akan ƙwayoyin halitta:
Bayanin SNP
Jiang S, Li S, Luo J, Wang X da Shi C (2023) Taswirar QTL da nazarin bayanai game da yawan sukari yayin nuna 'ya'yan itacePyrus pyrifolia.Gaba. Masana kimiyya na Shuke-shuke.14:1137104. doi: 10.3389/fpls.2023.1137104
Li, J., Zhang, Y., Ma, R., Huang, W., Hou, J., Fang, C., & Sun, L. (2022). Gano st1 yana nuna zaɓi wanda ya haɗa da tafiya a kan siffa da kuma yawan mai yayin kiwon waken soya.Mujallar Fasahar Halittu ta Shuke-shuke, 20(6), 1110-1121. https://doi.org/10.1111/pbi.13791
Xu, P., Zhang, X., Wang, X.da sauransu.Jerin kwayoyin halitta da bambancin kwayoyin halitta na kifin carp na yau da kullun,Cyprinus carpio.Nat Genet 46, 1212–1219 (2014). https://doi.org/10.1038/ng.3098
Zhuang, W., Chen, H., Yang, M.da sauransu.Tsarin halittar gyada da aka noma yana ba da haske game da karyotypes na legume, juyin halittar polyploid da kuma yadda ake kula da amfanin gona.Nat Genet 51, 865–876 (2019). https://doi.org/10.1038/s41588-019-0402-2
| Shekara | Mujallar Jarida | IF | Lakabi | Aikace-aikace |
| 2022 | Sadarwar Yanayi | 17.694 | Tushen kwayoyin halitta na giga-chromosomes da giga-genome na itacen peony Paeonia ostii | SLAF-GWAS |
| 2015 | Sabon Likitan Kwayoyin Halitta | 7.433 | Takaddun sawun gida sun mamaye yankunan kwayoyin halitta masu mahimmanci a fannin noma waken soya | SLAF-GWAS |
| 2022 | Mujallar Bincike Mai Ci Gaba | 12,822 | Shigar da Gossypium barbadense ta hanyar amfani da kwayoyin halitta a cikin G. hirsutum yana nuna kyakkyawan wuri don inganta ingancin auduga da yawan amfanin ƙasa a lokaci guda halaye | SLAF-Genetics na juyin halitta |
| 2019 | Masana'antar Kwayoyin Halitta | 10.81 | Binciken Halittar Jama'a da Majalisar De Novo Sun Bayyana Asalin Weedy Shinkafa a Matsayin Wasan Juyin Halitta | SLAF-Genetics na juyin halitta |
| 2019 | Halittar Halittar Halitta | 31.616 | Jerin kwayoyin halitta da bambancin kwayoyin halitta na kifi na yau da kullun, Cyprinus carpio | Taswirar Haɗin SLAF |
| 2014 | Halittar Halittar Halitta | 25.455 | Tsarin halittar gyada da aka noma yana ba da haske game da karyotypes na legume, polyploid juyin halitta da kuma kiwon amfanin gona. | Taswirar Haɗin SLAF |
| 2022 | Mujallar Fasahar Halittu ta Shuke-shuke | 9.803 | Gano ST1 ya nuna zaɓi wanda ya shafi hawan igiyar ruwa na siffar iri da kuma yawan mai a lokacin kiwon waken soya | Ci gaban Alamar SLAF |
| 2022 | Mujallar Duniya ta Kimiyyar Molecular | 6.208 | Ganowa da Ƙirƙirar Alamar DNA don Alkama-Leymus mollis 2Ns (2D) Sauyawar Chromosome ta Disomic | Ci gaban Alamar SLAF |
| Shekara | Mujallar Jarida | IF | Lakabi | Aikace-aikace |
| 2023 | Iyakoki a kimiyyar tsirrai | 6.735 | Taswirar QTL da nazarin bayanan sukari yayin nuna 'ya'yan itace na Pyrus pyrifolia | Taswirar Halitta |
| 2022 | Mujallar Fasahar Halittu ta Shuke-shuke | 8.154 | Gano ST1 ya nuna zaɓi wanda ya haɗa da hawan igiyar iri da kuma yawan mai a lokacin kiwon waken soya.
| Kiran SNP |
| 2022 | Iyakoki a kimiyyar tsirrai | 6.623 | Taswirar Haɗin Gwiwa na Halittu Masu Alaƙa da Hullless a Muhalli na Fari.
| GWAS |
