Non-Reference based mRNA Sequencing-NGS
Mawonekedwe
● Kujambula poly mRNA musanayambe kukonzekera laibulale
● Kusadalira ma genome aliwonse: kutengera zolembedwa za de novo, kutulutsa mndandanda wa ma unigene omwe ali ndi ma database angapo (NR, Swiss-Prot, COG, KOG, eggNOG, Pfam, GO, KEGG)
● Kusanthula kwatsatanetsatane kwa bioinformatic ya gene expression ndi transcript
Ubwino wa Utumiki
●Katswiri Wazambiri: Pokhala ndi mbiri yakukonza zitsanzo zopitilira 600,000 pa BMKGENE, zokhala ndi zitsanzo zamitundu yosiyanasiyana monga zikhalidwe zama cell, minyewa, ndi madzi amthupi, gulu lathu limabweretsa zambiri pantchito iliyonse. Tatseka bwino mapulojekiti opitilira 100,000 mRNA-Seq m'malo osiyanasiyana ofufuza.
●Kuwongolera Kwabwino Kwambiri: Timagwiritsa ntchito mfundo zowongolera pamagawo onse, kuyambira pakukonza zachitsanzo ndi laibulale mpaka kutsatizana ndi bioinformatics. Kuyang'anitsitsa mosamalitsa kumeneku kumatsimikizira kuperekedwa kwa zotsatira zabwino nthawi zonse.
● Mawu Omveka: timagwiritsa ntchito nkhokwe zingapo kuti tifotokozere za Differentially Expressed Genes (DEGs) ndikupanga kusanthula kofananirako, kupereka zidziwitso pama cell ndi ma cell omwe amatengera kuyankha kwa transcriptome.
●Thandizo la Post-Sales: Kudzipereka kwathu kumapitilira kutha kwa projekiti ndi nthawi ya miyezi itatu mutagulitsa. Panthawiyi, timapereka zotsatila za polojekiti, chithandizo chazovuta, ndi magawo a Q&A kuti tiyankhe mafunso aliwonse okhudzana ndi zotsatira.
Zitsanzo Zofunika ndi Kutumiza
| Library | Njira yotsatirira | Deta yovomerezeka | Kuwongolera Kwabwino |
| Poly A yowonjezera | Chithunzi cha PE150 Chithunzi cha DNBSEQ-T7 | 6-10 Gb | Q30≥85% |
Zitsanzo Zofunika:
Nucleotides:
| Conc.(ng/μl) | Mtengo (μg) | Chiyero | Umphumphu |
| ≥ 10 | ≥ 0.2 | OD260/280=1.7-2.5 OD260/230=0.5-2.5 Zochepa kapena zopanda mapuloteni kapena DNA kuipitsidwa komwe kumawonetsedwa pa gel. | Kwa zomera: RIN≥4.0; Kwa nyama: RIN≥4.5; 5.0≥28S/18S≥1.0; kukwera kochepa kapena kopanda koyambira |
● Zomera:
Muzu, tsinde kapena petal: 450 mg
Tsamba kapena Mbewu: 300 mg
Chipatso: 1.2 g
● Chinyama:
Mtima kapena matumbo: 300 mg
Viscera kapena Ubongo: 240 mg
Minofu: 450 mg
Mafupa, Tsitsi kapena Khungu: 1g
● Matenda a nyamakazi:
Tizilombo: 6g
Crustacea: 300 mg
● Mwazi wonse:1 chubu
● Maselo: 106 maselo
Kutumiza Kwachitsanzo Kovomerezeka
Chidebe: 2 ml centrifuge chubu (zojambula za malata ndizosavomerezeka)
Zitsanzo zolembera: Gulu+ lofanana mwachitsanzo A1, A2, A3; B1, B2, B3.
Kutumiza:
1. Madzi owuma: Zitsanzo ziyenera kupakidwa m'matumba ndikukwiriridwa mu ayezi wouma.
2. RNAstable chubu: RNA zitsanzo akhoza zouma mu RNA kukhazikika chubu (mwachitsanzo RNAstable®) ndi kutumizidwa mu firiji.
Kuyenda kwa Ntchito Yantchito
Kuyesera kupanga
Kupereka zitsanzo
Kusintha kwa RNA
Kumanga laibulale
Kutsata
Kusanthula deta
Pambuyo pogulitsa ntchito
Bioinformatics
Msonkhano wa Transcriptome ndi kusankha kwa unigene
Unigene annotation
Zitsanzo zogwirizana ndi kuwunika kwa zofananira zamoyo
Mitundu Yosiyanasiyana (DEGs)
Kufotokozera kwa magwiridwe antchito a DEGs
Kupititsa patsogolo ntchito kwa DEGs
Onani kupita patsogolo kotsogozedwa ndi BMKGene's eukaryotic NGS mRNA yotsatizana ntchito kudzera m'magulu osindikizidwa.
Shen, F. et al. (2020) 'De novo transcriptome assembly and sex-biased gene expression in the gonads of Amur catfish (Silurus asotus)', Genomics, 112 (3), pp. 2603-2614. doi: 10.1016/J.YGENO.2020.01.026.
Zhang, C. et al. (2016) 'Kusanthula kwa Transcriptome kwa sucrose metabolism panthawi ya kutupa kwa babu ndi chitukuko cha anyezi (Allium cepa L.)', Frontiers in Plant Science, 7 (September), p. 212763. doi: 10.3389/FPLS.2016.01425/BIBTEX.
Zhu, C. et al. (2017) 'De novo assembly, characterization and annotation for the transcriptome of Sarcocheilichthys sinensis', PLoS ONE, 12(2). doi: 10.1371/JOURNAL.PONE.0171966.
Zou, L. et al. (2021) 'De novo transcriptome analysis imapereka chidziwitso cha kulolerana kwa mchere wa Podocarpus macrophyllus pansi pa salinity stress', BMC Plant Biology, 21 (1), pp. 1-17. doi: 10.1186/S12870-021-03274-1/ZINTHU/9.
