Takamaiman-Locus Amplified Sequencing Sequencing (SLAF-Seq)
Gudun aiki
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.
Ana iya amfani da wannan fasaha tare da ko ba tare da wani nau'in kwayar halitta ba, tare da bututun bioinformatic daban-daban na kowane hali:
Tare da kwayoyin halitta: SNP da gano InDel
Ba tare da genome ba: tarin samfuri da gano SNP
● A cikincikin 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: Haɗa babban tsarin lambar lambar lamba biyu yana ba da damar yin jerin gwano na yawan jama'a a lokaci guda, kuma ƙayyadaddun ƙayyadaddun wuri yana haɓaka inganci, yana tabbatar da cewa lambobin tag ɗin 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. Thecikin silikipre-tsari ne da za'ayi don tabbatar da mafi kyau duka enzyme zane.
● Babban inganci a cikin narkewar Enzymatic: Gudanar da wanicikin silikiriga-kafi 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: Ƙungiyarmu tana kawo kwarewa mai yawa ga kowane aikin, tare da tarihin 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: BMKGENE ya 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 jerin 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
Ya haɗa da bincike mai zuwa:
- Bayanan Bayani na QC
- SLAF tag ci gaba
Yin taswira zuwa ga genome
Ba tare da kwayar halitta ba: tari
- Analysis of SLAF tags.: statistics, rarraba a fadin genome
- Gano alamar: SNP, InDel, SNV, kiran CV da annotation
Rarraba alamun SLAF akan chromosomes:
Rarraba SNP akan chromosomes:
Rahoton da aka ƙayyade na SNP
| 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 Biotechnology Journal | 9.803 | Gane ST1 yana bayyana zaɓin da ya haɗa da ƙwanƙwasa ƙwayoyin halittar iri da abun da ke cikin mai a lokacin gidan waken soya | 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 Biotechnology Journal | 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 |
