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Takamaiman-Locus Amplified Sequencing Sequencing (SLAF-Seq)

Takamaiman Wuraren Ƙarfafa Ƙarfafa Jeri (SLAF-Seq) Hoton da Aka Fitar

Wannan hanyar da BMKGene ta ɓullo da kanta, za'a iya rarraba ta a cikin rage yawan jeri na genome. Yana inganta ƙayyadaddun enzyme da aka saita don kowane aiki. Wannan yana tabbatar da samar da adadi mai yawa na alamun SLAF (yankin 400-500 bps na genome da ake jera su) waɗanda aka rarraba iri ɗaya a cikin kwayoyin halitta yayin da suke guje wa yankuna masu maimaitawa yadda ya kamata, don haka yana tabbatar da mafi kyawun gano alamun kwayoyin halitta.

Yana ba da saurin genotyping kuma yana kafa tushe don gano kwayoyin halitta mai aiki ko nazarin juyin halitta yana rage farashin kowane samfurin yayin da yake kiyaye inganci a cikin gano alamar kwayoyin halitta. RRGS yana samun wannan ta hanyar narkar da DNA tare da ƙuntatawa enzymes da kuma mai da hankali kan takamaiman girman girman guntu, ta haka ne ke tsara juzu'i na kwayoyin halitta. Daga cikin hanyoyin RRGS iri-iri, Specific-Locus Amplified Fragment Sequencing (SLAF) tsari ne na musamman da inganci.

Tuntube Mu

Cikakkun Sabis

Bioinformatics

Sakamakon Demo

Fitattun wallafe-wallafe

Gudun aiki

Sabis ɗin yana da wasu ƙirar riga-kafi a cikin siliki don ba da tabbacin zaɓin enzyme mafi kyau akan shirye-shiryen ɗakin karatu.

Tsarin Fasaha

Siffofin Sabis

● Yin aiki akan NovaSeq tare da PE150.

● Shirye-shiryen ɗakin karatu tare da barcoding sau biyu, yana ba da damar haɗawa sama da samfuran 1000.

● Mai zaman kansa ga kwayoyin halitta:

Tare da kwayoyin halitta: SNP da gano InDel

Ba tare da genome ba: tarin samfuri da gano SNP

● A cikinin-silikipre-tsari mataki mahara hani enzyme hadewa ana dubawa don nemo wadanda ke samar da daidaitaccen rarraba alamun SLAF tare da kwayoyin halitta.

● A lokacin gwaji na farko, an gwada haɗin enzyme guda uku a cikin samfurori na 3 don samar da ɗakunan karatu na SLAF na 9, kuma ana amfani da wannan bayanin don zaɓar haɗin haɗin enzyme mafi kyau don aikin.

Amfanin Sabis

●Babban Gano Alamar Halitta: Muna haɗa babban tsarin lambar lambar lamba biyu wanda ke ba da izinin yin jerin gwano na yawan jama'a a lokaci guda, da ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun bayanai, tabbatar da cewa lambobin alamar sun cika buƙatun daban-daban na tambayoyin bincike daban-daban.

● Low Dogara akan Genome: Ana iya amfani da shi ga nau'in tare da ko ba tare da alamar kwayar halitta ba.

●Zane mai sassauƙan tsari: Single-enzyme, dual-enzyme, multi-enzyme digestion, da nau'o'in enzymes daban-daban za a iya zaɓar su don cimma burin bincike daban-daban ko nau'in.

● Babban inganci a cikin narkewar Enzymatic: Gudanar da waniin-silikipre-tsari da kuma gwajin gwaji ya tabbatar da kyakkyawan tsari tare da ko da rarraba alamun SLAF akan chromosome (1 SLAF tag / 4Kb) da rage yawan maimaitawa (<5%).

●Ƙwararren Ƙwararru: Muna kawo kwarewa mai yawa ga kowane aiki, tare da rikodin rufewa fiye da 5000 SLAF-Seq ayyukan akan daruruwan nau'in, ciki har da tsire-tsire, dabbobi masu shayarwa, tsuntsaye, kwari, da kwayoyin ruwa.

● Gudun Aiki na Bioinformatic da kansa: Mun haɓaka aikin haɗin gwiwar bioinformatic mai haɗaka don SLAF-Seq don tabbatar da aminci da daidaito na fitowar ƙarshe.

Ƙayyadaddun Sabis

Nau'in bincike	Ma'aunin yawan jama'a da aka ba da shawarar	Dabarun jeri
		Zurfin jeri tag	Lambar tag
Taswirorin Halittu	2 iyaye da >150 zuriya	Iyaye: 20x WGS Saukewa: 10x	Girman kwayoyin halitta: <400 Mb: WGS ana ba da shawarar <1Gb: 100K tags 1-2Gb:: 200K tags > 2Gb: 300K tags Max 500k tags
Nazarin Ƙungiyar Genome-Wide (GWAS)	≥200 samfurori	10x ku
Juyin Halitta	≥30 samfurori, tare da> 10 samfurori daga kowane rukuni	10x ku

Bukatun Sabis

Ƙaddamarwa ≥ 5 ng/µL

Jimlar adadin ≥ 80 ng

Nandrop OD260/280=1.6-2.5

Agarose gel: a'a ko ƙayyadaddun ƙasƙanci ko gurɓatawa

Isar da Samfurin Nasiha

Kwantena: 2 ml centrifuge tube

(Ga mafi yawan samfurori, muna ba da shawarar kada a adana a cikin ethanol)

Alamar samfurin: Samfuran suna buƙatar a yi musu lakabi a sarari kuma iri ɗaya ga siffan bayanin samfurin da aka ƙaddamar.

Jirgin ruwa: Busasshen ƙanƙara: Ana buƙatar a haɗa samfuran a cikin jakunkuna da farko kuma a binne su a bushe-kankara.

Gudun Aikin Sabis

Misalin QC

Gwajin matukin jirgi

SLAF-gwaji

Shirye-shiryen Laburare

Jeri

Binciken Bayanai

Bayan-tallace-tallace Services

Na baya: Tsarin DNA/RNA - Mai bin Nanopore

Na gaba: Tsarin Dan Adam Gabaɗaya Exome

Binciken mu na bioinformatical ya ƙunshi:

Data QC da datsa bayanai don cire N-rich reads, adaftan karantawa ko ƙarancin karantawa.

Kula da inganci na biyu na mai tsabta yana karantawa don bincika rarraba tushe, ƙimar jeri da ƙididdigar bayanai, amma kuma don bincika ingantaccen narkewa da abubuwan da aka samu.

Da zarar an duba karatun, akwai zaɓuɓɓuka biyu:

Yin taswira zuwa ga genome
Ba tare da kwayar halitta ba: tari

Bayan shi, ana amfani da nazarin alamun SLAF don yin wasu kira daban-daban don taimakawa tare da gano alamar: SNP, InDel, SNV, kiran CV da annotation.

Rarraba alamun SLAF akan chromosomes:

Rarraba SNP akan chromosomes:

Rahoton da aka ƙayyade na SNP

Jiang S, Li S, Luo J.Pyrus pyrifolia.Gaba. Shuka Sci.14:1137104. doi: 10.3389/fpls.2023.1137104

Li, J., Zhang, Y., Ma, R., Huang, W., Hou, J., Fang, C., & Sun, L. (2022). Gane st1 yana bayyana zaɓin da ya haɗa da ƙwanƙwasa nau'in halittar iri da abun cikin mai yayin cikin gida na waken soya.Jaridar Kimiyyar Kimiyyar Tsirrai, 20 (6), 1110-1121. doi.org/10.1111/pbi.13791

Xu, P., Zhang, X., Wang, X.da al.Tsarin kwayoyin halitta da bambancin jinsin carp na kowa,Cyprinus carpio.Ina Genet 46, 1212-1219 (2014). https://doi.org/10.1038/ng.3098

Zhuang, W., Chen, H., Yang, M.da al.Halin halittar gyada da aka noma yana ba da haske game da legume karyotypes, juyin halittar polyploid da amfanin gona na gida.Ina Genet 51, 865-876 (2019). doi.org/10.1038/s41588-019-0402-2

Shekara	Jarida	IF	Take	Aikace-aikace
2022	Sadarwar yanayi	17.694	Tushen genomic na giga-chromosomes da giga-genome na itacen peony Paeonia ostii	SLF-GWAS
2015	Sabon Likitan Halitta	7.433	Sawun cikin gida yana kafa yankuna masu mahimmancin agronomic a cikin waken soya	SLF-GWAS
2022	Jaridar Babban Bincike	12.822	G. hirsutum na Gossypium barbadense mai faɗin ɗan adam bayyana loci mafi girma don haɓaka ingancin fiber auduga da yawan amfanin ƙasa lokaci guda halaye	SLAF-Genetics na Juyin Halitta
2019	Shuka Kwayoyin Halitta	10.81	Binciken Al'umma na Al'umma da Majalisar De Novo sun Bayyana Asalin Weedy Shinkafa a matsayin Wasan Juyin Halitta	SLAF-Genetics na Juyin Halitta
2019	Halitta Halitta	31.616	Tsarin kwayoyin halitta da bambancin kwayoyin halittar irin kifi na kowa, Cyprinus carpio	SLAF-Haɗin taswira
2014	Halitta Halitta	25.455	Halin halittar gyada da aka noma yana ba da haske game da legume karyotypes, polyploid juyin halitta da amfanin gona domestication.	SLAF-Haɗin taswira
2022	Jaridar Kimiyyar Kimiyyar Tsirrai	9.803	Gane ST1 yana bayyana zaɓin da ya haɗa da ƙwanƙwasa ƙwayoyin halittar iri da abun da ke ciki na mai a lokacin gidan waken suya	SLF-Marker ci gaba
2022	Jaridar Duniya ta Kimiyyar Kwayoyin Halitta	6.208	Ganewa da Ci gaban Alamar DNA don Alkama-Leymus mollis 2Ns (2D) Sauya Disomic Chromosome	SLF-Marker ci gaba

Shekara	Jarida	IF	Take	Aikace-aikace
2023	Gaba a kimiyyar shuka	6.735	Taswirar QTL da bincike na kwafi na abun ciki na sukari yayin girbin 'ya'yan itace na Pyrus pyrifolia	Taswirar Halittu
2022	Jaridar Kimiyyar Kimiyyar Tsirrai	8.154	Gano ST1 yana bayyana zaɓin da ya haɗa da ƙwanƙwasa nau'in ilimin halittar iri da abun cikin mai yayin cikin gida na waken soya.	Farashin SNP
2022	Gaba a kimiyyar shuka	6.623	Taswirar Ƙungiya mai Faɗin Halitta na Hulless Barely Phenotypes a cikin Muhallin Fari.	GWAS