usoro nke circRNA-Ilumina
Atụmatụ
● Mbelata rRNA na-esochi nkwadebe ọbá akwụkwọ ntụzịaka, na-eme ka data usoro nke akọwapụtara nke ọma.
● Bioinformatic workflow na-enyere aka ịkọ amụma circRNA na ọnụọgụ okwu
Uru ọrụ
●Ọbá akwụkwọ RNA zuru oke:anyị na-eji nbibi rRNA kama ịbelata RNA linear na nkwadebe tupu ụlọ akwụkwọ anyị, na-ahụ na data usoro nke gụnyere ọ bụghị naanị circRNA kamakwa mRNA na lncRNA, na-eme ka nyocha njikọ dị na datasets ndị a.
●Nyocha nhọrọ nke netwọk RNA endogenous asọmpi (ceRNA).: inye nghọta miri emi n'ime usoro nhazi ekwentị
●Ọkachamara sara mbara: na ndekọ ndekọ nke nhazi ihe karịrị 20,000 samples na BMK, na-agbasa ụdị ụdị dị iche iche na ọrụ lncRNA, ndị otu anyị na-eweta ahụmahụ 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.
● 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 zaa ajụjụ ọ bụla metụtara nsonaazụ ya.
Ihe nlele chọrọ na nnyefe
| Ụlọ akwụkwọ | Platform | Data akwadoro | Data QC |
| Poly A bara ụba | Ilumina PE150 | 16-20 Gb | Q30≥85% |
Ihe atụ chọrọ:
Nucleotides:
| Conc.(ng/μl) | Ọnụ ego (μg) | Ịdị ọcha | Iguzosi ike n'ezi ihe |
| ≥ 100 | ≥ 0.5 | 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. | Maka osisi: RIN≥6.5; Maka anụmanụ: RIN≥7.0; 5.0≥28S/18S≥1.0; nwere oke ma ọ bụ enweghị mgbago elu |
● Osisi:
Mgbọrọgwụ, azuokokoosisi ma ọ bụ petal: 450 mg
Akwụkwọ ma ọ bụ mkpụrụ: 300 mg
Mkpụrụ: 1.2 g
● Anụmanụ:
Obi ma ọ bụ eriri afọ: 450 mg
Viscera ma ọ bụ ụbụrụ: 240 mg
Akwara: 600 mg
Ọkpụkpụ, ntutu ma ọ bụ akpụkpọ: 1.5g
● Arthropods:
Ụmụ ahụhụ: 9g
Crustacea: 450 mg
● Ọbara dum:2 ọkpọkọ
● Selụ: 106 mkpụrụ ndụ
● Serum na Plasma: 6 ml
Nnyefe nlele akwadoro
Akpa: 2 ml centrifuge tube (Tin foil adịghị akwadoro)
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 ice-akọrọ.
2. tubes RNAstable: Enwere ike ịkpọsa ihe nlele RNA na tube stabilization RNA (dịka ọmụmaatụ RNAstable®) ma bufee ya n'ime ụlọ okpomọkụ.
Usoro ọrụ ọrụ
Nnwale imewe
Nbufe ihe atụ
mmịpụta RNA
Ịrụ ụlọ akwụkwọ
Usoro
Nyocha data
Mgbe-ire ọrụ
Bioinformatics
amụma circRNA: nkesa chromosomal
CirRNA ndị egosipụtara n'ụzọ dị iche - nkata mgbawa ugwu
CirRNA ndị egosipụtara n'ụzọ dị iche - nchịkọta usoro nhazi
Ọgaranya na-arụ ọrụ nke mkpụrụ ndụ ihe nketa circRNA
Nyochaa ọganihu nyocha nke ọrụ nhazi usoro BMKGene' circRNA kwadoro site na nchịkọta mbipụta.
Wang, X. et al.(2021) 'CPSF4 na-ahazi nhazi circRNA na microRNA na-agbachitere mkpụrụ ndụ ihe nketa n'ime carcinoma hepatocellular', Oncogene 2021 40:25, 40(25), p. 4338-4351.doi: 10.1038/s41388-021-01867-6.
Xia, K. et al.(2023) 'X oo-responsive transcriptome na-ekpughe ọrụ okirikiri RNA133 na nguzogide ọrịa site n'ịchịkwa okwu OsARAB na osikapa', Nnyocha Phytopathology, 5(1), p. 1-14.doi: 10.1186/S42483-023-00188-8/FIGURES/6.
Y, H. et al.(2023) 'CPSF3 na-agbanwe nguzozi nke ederede okirikiri na ahịrị na carcinoma hepatocellular.doi: 10.21203/RS.3.RS-2418311/V1.
Zhang, Y. et al.(2023) 'Nnyocha zuru oke nke circRNAs na cirrhotic cardiomyopathy tupu na mgbe transplantation imeju', International Immunopharmacology, 114, p.109495. doi: 10.1016/J.INTIMP.2022.109495.
