Usoro transcriptome niile - Illumina
Atụmatụ
● Ọbá akwụkwọ abụọ iji hazie transcriptome zuru ezu: mmebi rRNA na-esote nkwadebe ọba akwụkwọ PE150 na nhọrọ nha na-esote nkwadebe ọba akwụkwọ SE50.
● Nyocha bioinformatics zuru ezu nke mRNA, lncRNA, circRNA na miRNA na akụkọ bioinformatics dị iche iche.
● Ntụle njikọta nke okwu RNA niile na akụkọ jikọtara ọnụ, gụnyere nyocha netwọkụ ceRNA.
Uru ọrụ
●Nyocha miri emi nke netwọk nchịkwa: A na-akwado nyocha netwọkụ ceRNA site na nhazi njikọ nke mRNA, lncRNA, circRNA na miRNA yana site na usoro ọrụ bioinformatic na-agwụ ike.
●Nkọwa zuru oke: anyị na-eji ọtụtụ ọdụ data na-arụ ọrụ ịkọwapụta Genes dị iche iche (DEGs) na-arụ ọrụ ma na-eme nyocha nke ịba ụba kwekọrọ, na-enye nghọta na usoro cellular na molecular na-akpata nzaghachi transcriptome.
●Ọkachamara sara mbara: na ndekọ egwu nke imechi ihe karịrị 2000 ọrụ transcriptome nke ọma na ngalaba nyocha dị iche iche, ndị otu anyị na-eweta ahụmịhe bara ụba na ọrụ ọ bụla.
●Njikwa ogo siri ike: anyị na-emejuputa isi ihe nchịkwa n'ofe niile, site na nlele na nkwadebe ọbá akwụkwọ ruo usoro na bioinformatics.Nleba anya nke ọma a na-eme ka o doo anya na nnyefe nke nsonaazụ dị elu mgbe niile.
● Nkọwa zuru oke: anyị na-eji ọtụtụ ọdụ data na-arụ ọrụ ịkọwapụta Genes dị iche iche (DEGs) na-arụ ọrụ ma na-eme nyocha nke ịba ụba kwekọrọ, na-enye nghọta na usoro cellular na molecular na-akpata nzaghachi transcriptome.
●Nkwado maka ire ere: Nkwenye anyị na-agafe karịa mmecha ọrụ yana oge ọrụ ọnwa 3 gachara.N'ime oge a, anyị na-enye nlebanya ọrụ, enyemaka nchọpụta nsogbu, yana oge ajụjụ & A iji lebara ajụjụ ọ bụla metụtara nsonaazụ ya.
Ihe nlele chọrọ na nnyefe
| Ụlọ akwụkwọ | Usoro usoro | akwadoro data | Njikwa ogo |
| rRNA agwụla | Ilumina PE150 | 16 gb | Q30≥85% |
| ahọpụtara nha | Illumina SE50 | 10-20M na-agụ |
Ihe atụ chọrọ:
Nucleotides:
| Conc.(ng/μl) | Ọnụ ego (μg) | Ịdị ọcha | Iguzosi ike n'ezi ihe |
| ≥ 100 | ≥ 1 | OD260/280=1.7-2.5 OD260/230=0.5-2.5 Enwere oke ma ọ bụ enweghị protein ma ọ bụ mmetọ DNA nke egosiri na gel. | Osisi: RIN≥6.5 Anụmanụ: RIN≥7.0 5.0≥28S/18S≥1.0; nwere oke ma ọ bụ enweghị mgbago elu |
Nnyefe nlele akwadoro
akpa:
2 ml nke centrifuge tube (anaghị akwado foil tin)
Ịkpọ aha nlele: Otu+ megharịa dịka A1, A2, A3;B1, B2, B3......
Mbupu:
1.Dry-ice: A ga-etinye ihe nlele n'ime akpa ma lie ya na akọrọ-ice.
2.RNAstable tubes: Enwere ike ihichapụ ihe nlele RNA na tube stabilization RNA (dịka RNAstable®) ma bufee ya na okpomọkụ.
Usoro ọrụ ọrụ
Nnwale imewe
Nbufe ihe atụ
mmịpụta RNA
Ịrụ ụlọ akwụkwọ
Usoro
Nyocha data
Mgbe-ire ọrụ
Bioinformatics
Nkọwa okwu RNA
Mkpụrụ ndụ ihe nketa egosipụtara dị iche iche
nyocha ceRNA
MiRNA ndị egosipụtara n'ụzọ dị iche na RNA ndị metụtara ya
Nyochaa ọganihu nyocha nke BMKGene' na-akwado ọrụ usoro transcriptome niile site na nchịkọta mbipụta.
Dai, Y. et al.(2022) ' Profaịlụ okwu zuru oke nke mRNAs, lncRNAs na miRNAs na ọrịa Kashin-Beck nke RNA-sequencing chọpụtara', Molecular Omics, 18(2), p. 154-166.doi: 10.1039/D1MO00370D.
Liu, N. nan et al.(2022) 'Nnyocha transcriptomes n'ogologo zuru oke nke nguzogide oyi nke Apis cerana na ugwu Changbai n'oge oke oyi.', Gene, 830, p. 146503–146503.doi: 10.1016/J.GENE.2022.146503.
Wang, XJ, et al.(2022) 'Multi-Omics Integration-based Prioritization of Enogenous RNA Regulation Networks in Small Cell Lung Cancer: Molecular Characteristics and Drug Candidates', Frontiers in Oncology, 12, p.904865. doi: 10.3389/FONC.2022.904865/BIBTEX.
Xu, P. et al.(2022) 'Ntụle agbakwunyere nke profaịlụ okwu lncRNA/circRNA-miRNA-mRNA na-ekpughe nghọta ọhụrụ banyere usoro nwere ike na nzaghachi nematodes mgbọrọgwụ na ahụekere', BMC Genomics, 23(1), p. 1-12.doi: 10.1186/S12864-022-08470-3/ỌNỤTA/7.
Yan, Z. et al.(2022) 'Nhazi usoro RNA zuru oke na-akọwapụta usoro mkpụrụ ndụ jikọtara ya na idobe ogo mkpụrụ akwụkwọ broccoli site na irradiation LED LED', Biology Postharvest and Technology, 188, p.111878. doi: 10.1016/J.POSTHARVBIO.2022.111878.
