Usoro nke Eukaryotic mRNA-NGS
Atụmatụ
● Njide poly mRNA tupu akwadebe ụlọ akwụkwọ
● Nkwadebe ọbá akwụkwọ mRNA ntụziaka nhọrọ iji mee ka ị nweta data nhazi usoro strand akọwapụtara
● Nyocha bioinformatic nke okwu mkpụrụ ndụ ihe nketa na nhazi ederede
Uru
●Ọkachamara sara mbara: Ndị otu anyị ahazila ihe karịrị 600,000 sample na BMKgene, na-agbasa ụdị ụdị dị iche iche dị ka omenala cell, anụ ahụ, na mmiri ahụ. Anyị na-eweta ahụmịhe bara ụba na ọrụ ọ bụla wee rụchaa ihe karịrị 100,000 mRNA-Seq nke ọma na ngalaba nyocha dị iche iche.
●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 ị nweta nsonaazụ dị elu mgbe niile, na-emesi gị obi ike na ọrụ gị dị n'aka ndị a pụrụ ịdabere na ya.
●Nkọwa zuru oke: Anyị na-eji ọtụtụ ọdụ data iji kọwapụta mkpụrụ ndụ ihe dị iche iche (DEGs) na-arụ ọrụ ma mee nyocha nkwalite nkwalite kwekọrọ. Ụzọ a sara mbara na-enye nghọta n'ime usoro cellular na molekụla n'okpuru nzaghachi transcriptome, na-eme ka ị mara nke ọma maka nsonaazụ ọrụ gị.
●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 | Usoro usoro | akwadoro data | Njikwa ogo |
| Poly A bara ụba | Ilumina PE150 DNBSEQ-T7 | 6-10 Gb | Q30≥85% |
Ihe atụ chọrọ:
Nucleotides:
| Conc.(ng/μl) | Ọnụ ego (μg) | Ịdị ọcha | Iguzosi ike n'ezi ihe | |
| Ọbá akwụkwọ ọkọlọtọ | ≥ 10 | ≥ 0.2 | 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≥4.0; Maka anụmanụ: RIN≥4.5; 5.0≥28S/18S≥1.0; nwere oke ma ọ bụ enweghị mgbago elu |
| Akwụkwọ ntuziaka | ≥ 10 | ≥ 0.2 | 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≥4.0; Maka anụmanụ: RIN≥4.5; 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: 300 mg
Viscera ma ọ bụ ụbụrụ: 240 mg
Akwara: 450 mg
Ọkpụkpụ, ntutu ma ọ bụ akpụkpọ: 1g
● Arthropods:
Ụmụ ahụhụ: 6g
Crustacea: 300 mg
● Ọbara dum: 1 tube
● Selụ: 106 mkpụrụ ndụ
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. RNA okpokoro tubes: Enwere ike ịkpọsa ihe nlele RNA na tube stabilization RNA (dịka ọmụmaatụ 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
Eukaryotic mRNA usoro nyocha usoro ọrụ
Bioinformatics
ØNjikwa ogo data raw
ØNtụaka genome alignment
ØNyocha nhazi nke transcript
ØNgosipụta okwu
ØNyocha okwu dị iche iche
ØNkọwa ọrụ na ịba ụba
Ntụaka genome alignment
Saturation data
Ihe nlere anya na ntule nke ntugharị ndu
Mkpụrụ ndụ ihe nketa egosipụtara dị iche iche (DEGs)
Nkọwa arụmọrụ nke DEGs
Ọganihu arụ ọrụ nke DEGs
Nyochaa ọganihu ndị ọrụ nhazi usoro eukaryotic NGS mRNA BMKGene na-akwado site na nchịkọta mbipụta.
Huang, L. et al. (2023) 'Triclosan na triclocarban na-eme ka ike ísì ụtọ nke azụ azụ azụ na-eme ka ike gwụ site n'ime ka ọ ghara ịdị na-esi ísì ụtọ, na-akpaghasị ntụgharị nrịanrịa, na nhazi ozi na-enye nsogbu', Nnyocha mmiri, 233, p. 119736. doi: 10.1016/J.WATRES.2023.119736.
Jia, LJ et al. (2023) 'Aspergillus fumigatus na-adọpụ mmadụ p11 iji redirect fungal-nwere phagosomes na ụzọ ndị na-adịghị emebi emebi', Cell Host & Microbe, 31 (3), p. 373-388.e10. doi: 10.1016/J.CHOM.2023.02.002.
Jin, K. et al. (2022) 'Ihe odide ederede TCP metụtara na mmepe nke Ma Bamboo (Dendrocalamus latiflorus Munro)', Frontiers in Plant Science, 13, p. 884443. doi: 10.3389/FPLS.2022.884443/BIBTEX.
Wen, X. et al. (2022) 'The Chrysanthemum lavandulifolium genome na usoro molekụla n'okpuru ụdị capitulum dị iche iche', Horticulture Research, 9. doi: 10.1093/HR/UHAB022.
Zhang, Yujie et al. (2023) 'A cascade nanoreactor maka ịkwalite ọgwụgwọ sonodynamic na ọrịa cancer colorectal site na synergistic ROS augment na autophagy blockage', Nano Today, 49, p. 101798. doi: 10.1016/J.NANTOD.2023.101798.
