Kutsata kwa Prokaryotic RNA
Mawonekedwe
● Kukonzekera kwachitsanzo kwa RNA kumaphatikizapo kuchepetsedwa kwa rRNA kutsatiridwa ndi kukonzekera kwa laibulale ya RNA.
● Kusanthula kwa bioinformatics kutengera kutsata kwa ma genome
● Kusanthula kumaphatikizapo mafotokozedwe a jini ndi ma DEG komanso mawonekedwe a zolemba ndi kusanthula kwa sRNA
Ubwino wa Utumiki
●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.
●Strand-specific Sequencing Data: chifukwa chakukonzekera laibulale ya RNA kukhala yolunjika, kumathandizira kuzindikira zolembedwa zotsutsana ndi malingaliro.
●Kusanthula Kwathunthu Kogwirizana ndi Prokaryotic Transcriptomes: payipi ya bioinformamatic imaphatikizapo osati kusanthula kwa mawu a jini komanso kusanthula kapangidwe kake, kuphatikiza kuzindikiritsa ma opereni, ma UTR ndi olimbikitsa. Zimaphatikizansopo kusanthula kwa ma sRNA, kutanthauza zofotokozera ndi kulosera za kapangidwe kachiwiri ndi zolinga.
●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 |
| rRNA yatha laibulale yowongolera | Chithunzi cha PE150 | 1-2 Gb | Q30≥85% |
Zitsanzo Zofunika:
| Conc.(ng/μl) | Mtengo (μg) | Chiyero | Umphumphu |
| ≥ 50 | ≥ 1 | OD260/280=1.8-2.0 OD260/230=1.0-2.5 Zochepa kapena zopanda mapuloteni kapena DNA kuipitsidwa komwe kumawonetsedwa pa gel. | RIN≥6.5 |
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
Kupereka zitsanzo
Kumanga laibulale
Kutsata
Kusanthula deta
Pambuyo pogulitsa ntchito
Bioinformamatic Analysis Workflow
Mulinso Kusanthula Kotere:
● Kuwongolera khalidwe la data yaiwisi
● Kuyanjanitsa ku genome
● Kuwunika Ubwino wa Library: Kugawikana kwa RNA mwachisawawa, kukula kwake ndi kachulukidwe kakutsatizana.
● Kafotokozedwe kake ka jini yoloseredwa
● Kusanthula Kufotokozera: Kugwirizana ndi Kusanthula Kwachinthu Chachikulu (PCA)
● Differential Gene Expression (DEGs)
● Ndemanga zogwira ntchito ndi kupititsa patsogolo ma DEG
● Kusanthula kwa sRNA: kulosera, kutanthauzira, chandamale, ndi kuneneratu kwachiwiri
● Transcript Structure Analysis: operons, kuyamba ndi kutsiriza maudindo, Untranslated dera (UTS), wolimbikitsa, ndi SNP/InDel kusanthula
Sequencing machulukitsidwe
Kafotokozedwe ka ntchito ya ma jini olembera
Kugwirizana pakati pa zitsanzo
Differential Expressed Genes (DEGs) kusanthula
Kusanthula kwa magwiridwe antchito
sRNA chidziwitso
Onani kupita patsogolo kotsogozedwa ndi BMKGene's Nanopore yotsatizanatsatizana za mRNA m'bukuli.
Guan, CP ndi al. (2018) 'Global Transcriptome Changes of Biofilm-Forming Staphylococcus epidermidis Responding to Total Alkaloids of Sophorea alopecuroides',Polish Journal ya Microbiology, 67(2), p. 223. doi: 10.21307/PJM-2018-024.
