Matsakaicin tushen mRNA Sequencing-NGS
Siffofin
● Ɗauki poly mRNA kafin shirya ɗakin karatu
● Mai zaman kansa ga kowane nau'in kwayar halitta: bisa de novo taro na kwafi, samar da jerin unigenes waɗanda aka annotate tare da mahara bayanai (NR, Swiss-Prot, COG, KOG, eggNOG, Pfam, GO, KEGG)
● Cikakken bincike na bioinformatic na maganganun kwayoyin halitta da tsarin kwafi
Amfanin Sabis
●Ƙwararren Ƙwararru: tare da rikodin waƙa na sarrafa samfuran sama da 600,000 a BMKGENE, wanda ke tattare da nau'ikan samfura daban-daban kamar al'adun tantanin halitta, kyallen takarda, da ruwan jiki, ƙungiyarmu tana kawo ƙwarewar ƙwarewa ga kowane aiki. Mun sami nasarar rufe ayyukan mRNA-Seq sama da 100,000 a fannonin bincike daban-daban.
●Tsananin Kula da Inganci: muna aiwatar da mahimman abubuwan sarrafawa a duk matakai, daga samfurin da shirye-shiryen ɗakin karatu zuwa jerin abubuwa da bioinformatics. Wannan saka idanu mai kyau yana tabbatar da isar da sakamako mai inganci akai-akai.
● Cikakken Bayani: muna amfani da ma'ajin bayanai da yawa don yin bayanin aikin Halittu Masu Bambance-bambance (DEGs) da aiwatar da ƙididdigar haɓaka daidai, samar da fahimta kan tsarin salon salula da ƙwayoyin cuta waɗanda ke ƙarƙashin amsawar kwafin.
●Tallafin Bayan Talla: alkawarinmu ya wuce bayan kammala aikin tare da lokacin sabis na watanni 3 bayan-sayar. A wannan lokacin, muna ba da bibiyar aikin, taimako na warware matsala, da kuma zaman Q&A don magance duk wata tambaya da ta shafi sakamakon.
Samfuran Bukatun da Bayarwa
| Laburare | Dabarun jeri | An ba da shawarar bayanai | Kula da inganci |
| Poly A ya wadata | Farashin PE150 DNBSEQ-T7 | 6-10 GB | Q30≥85% |
Samfuran Bukatun:
Nucleotides:
| Conc.(ng/μl) | Adadin (μg) | Tsafta | Mutunci |
| ≥ 10 | 0.2 | OD260/280=1.7-2.5 OD260/230=0.5-2.5 Iyakance ko babu furotin ko gurɓataccen DNA da aka nuna akan gel. | Don tsire-tsire: RIN≥4.0; Na dabbobi: RIN≥4.5; 5.0≥28S/18S≥1.0; iyakance ko babu girma daga tushe |
● Shuke-shuke:
Tushen, kara ko petal: 450 MG
Leaf ko iri: 300 MG
'Ya'yan itace: 1.2 g
● Dabbobi:
Zuciya ko hanji: 300 MG
Viscera ko Brain: 240 MG
tsoka: 450 MG
Kasusuwa, Gashi ko Fata: 1g
● Arthropods:
Kwari: 6g
Crustacea: 300 MG
● Jini dukaku: 1 tube
● Kwayoyin: 106 Kwayoyin
Isar da Samfurin Nasiha
Kwantena: 2 ml bututu centrifuge (Ba a ba da shawarar foil tin ba)
Alamar samfur: Ƙungiya+ kwafi misali A1, A2, A3; B1, B2, B3.
Kawo:
1. Busasshen ƙanƙara: Ana buƙatar samfurori a cikin jaka kuma a binne su a bushe-kankara.
2. RNAstable tubes: Ana iya bushe samfuran RNA a cikin bututun daidaitawar RNA (misali RNAstable®) kuma a tura su cikin zafin jiki.
Gudun Aikin Sabis
Gwajin ƙira
Samfurin bayarwa
RNA cirewa
Gina ɗakin karatu
Jeri
Binciken bayanai
Bayan-sayar da sabis
Bioinformatics
Taro rubutun taro da zaɓin unigene
Bayanin Unigene
Samfurin daidaitawa da ƙima na kwafin halittu
Ƙwayoyin Halittu Masu Bambance-bambance (DEGs)
Bayanin Aiki na DEGs
Haɓaka Aiki na DEGs
Bincika ci gaban da BMKGene's eukaryotic NGS mRNA sequencing sabis na eukaryotic ta hanyar tarin wallafe-wallafe.
Shen, F. et al. (2020) 'De novo transcriptome taro da jinsin son zuciya magana a cikin gonads na Amur catfish (Silurus asotus)', Genomics, 112(3), shafi. 2603-2614. doi: 10.1016/J.YGENO.2020.01.026.
Zhang, C. et al. (2016) 'Transcriptome bincike na sucrose metabolism a lokacin kwan fitila kumburi da ci gaba a cikin albasa (Allium cepa L.)', Frontiers in Plant Science, 7 (Satumba), p. 212763. doi: 10.3389/FPLS.2016.01425/BIBTEX.
Zhu, C. et al. (2017) 'De novo taro, halayyar da annotation ga fassarar Sarcocheilichthys sinensis', PLoS ONE, 12 (2). doi: 10.1371/JOURNAL.PONE.0171966.
Zo, L. et al. (2021) 'De novo transcriptome bincike yana ba da haske game da haƙurin gishiri na Podocarpus macrophyllus a ƙarƙashin damuwa salinity', BMC Plant Biology, 21(1), shafi 1-17. doi: 10.1186/S12870-021-03274-1/FIGURES/9.
