circRNA sequencing-Illumina
Mawonekedwe
● Kuwonongeka kwa rRNA kutsatiridwa ndi kukonzekera laibulale yotsogolera, kuthandizira kutsatizana kwa deta yomwe ili yokhazikika.
● Bioinformamatic workflow imathandizira kulosera kwa circRNA ndi kuchuluka kwa mawu
Ubwino wa Utumiki
●Ma library ambiri a RNA:timagwiritsa ntchito kuchepa kwa rRNA m'malo mwa kutha kwa mzere wa RNA pokonzekera laibulale yathu isanayambe, kuonetsetsa kuti zotsatizana zikuphatikiza osati circRNA komanso mRNA ndi lncRNA, zomwe zimathandizira kusanthula kophatikizana pamagulu awa.
●Kusanthula kosankha kwa maukonde amtundu wa RNA (ceRNA) wampikisano: Kupereka zidziwitso zakuya pamakina oyendetsera ma cell
●Katswiri Wazambiri: yokhala ndi mbiri yakukonza zitsanzo zopitilira 20,000 ku BMK, zokhala ndi zitsanzo zamitundu yosiyanasiyana ndi mapulojekiti a lncRNA, gulu lathu limabweretsa zokumana nazo zambiri pantchito iliyonse.
●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.
● Thandizo Pambuyo pa Kugulitsa: 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 | nsanja | Deta yovomerezeka | Zithunzi za QC |
| Poly A yowonjezera | Chithunzi cha PE150 | 16-20 Gb | Q30≥85% |
Zitsanzo Zofunika:
Nucleotides:
| Conc.(ng/μl) | Mtengo (μg) | Chiyero | Umphumphu |
| ≥100 | ≥ 0.5 | 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≥6.5; Kwa nyama: RIN≥7.0; 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: 450 mg
Viscera kapena Ubongo: 240 mg
Minofu: 600 mg
Mafupa, Tsitsi kapena Khungu: 1.5g
● Matenda a nyamakazi:
Kulemera kwake: 9g
Krustacea: 450 mg
● Magazi Onse:2 machubu
● Maselo: 106 maselo
● Seramu ndi Plasmapa: 6ml
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. Dry-ice: 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
Kuneneratu kwa circRNA: kugawa kwa chromosomal
Ma circRNA Osiyanasiyana - Chiwembu cha Volcano
Ma circRNA Owonetsedwa Mosiyana - magulu otsogola
Kupititsa patsogolo magwiridwe antchito amtundu wa circRNA's host
Onani kupita patsogolo kwa kafukufuku woyendetsedwa ndi BMKGene 'circRNA sequencing services kudzera m'magulu osindikizidwa.
Wang, X. et al.(2021) 'CPSF4 imayendetsa mapangidwe a circRNA ndi microRNA mediated gene silence mu hepatocellular carcinoma', Oncogene 2021 40:25, 40 (25), pp. 4338-4351.doi: 10.1038/s41388-021-01867-6.
Xia, K. et al.(2023) 'X oo-responsive transcriptome imasonyeza ntchito ya RNA133 yozungulira polimbana ndi matenda mwa kuwongolera mawu a OsARAB mu mpunga', Phytopathology Research, 5 (1), pp. 1-14.doi: 10.1186/S42483-023-00188-8/ZINTHU/6.
Y, H. et al.(2023) 'CPSF3 imasintha zolembedwa zozungulira komanso zozungulira mu hepatocellular carcinoma'.doi: 10.21203/RS.3.RS-2418311/V1.
Zhang, Y. et al.(2023) 'Kuwunika kwathunthu kwa circRNAs mu cirrhotic cardiomyopathy isanachitike komanso pambuyo pa kupatsirana kwa chiwindi', International Immunopharmacology, 114, p.109495. doi: 10.1016/J.INTIMP.2022.109495.
