ʻO ke kaʻina transcriptome holoʻokoʻa - Illumina
Nā hiʻohiʻona
● Hale waihona lua e hoʻonohonoho i ka transcriptome holoʻokoʻa: hoʻopau ʻia ka rRNA a ukali ʻia e ka hoʻomākaukau hale waihona PE150 a me ke koho nui a ukali ʻia e ka hoʻomākaukau waihona SE50
● Hoʻopiha piha i ka bioinformatics ka nānā ʻana o mRNA, lncRNA, circRNA a me miRNA i nā hōʻike bioinformatics ʻokoʻa
● Ka nānā 'ana o nā hua'ōlelo RNA a pau ma ka hō'ike hui pū, me ka nānā 'ana i nā pūnaewele ceRNA.
Pono lawelawe
●ʻIke hohonu o nā pūnaewele hoʻoponopono: Hoʻohana ʻia ka ʻikena pūnaewele ceRNA e ka hui pū ʻana o ka mRNA, lncRNA, circRNA a me miRNA a me kahi kahe hana bioinformatic pau loa.
●Hōʻikeʻike piha: hoʻohana mākou i nā ʻikepili he nui e hoʻopuka pono i nā Genes Differentially Expressed (DEGs) a hana i ka loiloi hoʻonui e pili ana, e hāʻawi ana i nā ʻike e pili ana i nā kaʻina kelepona a me nā molekala e pili ana i ka pane transcriptome.
●ʻIke Nui: me kahi moʻolelo o ka hoʻopau kūleʻa ʻana ma luna o 2000 mau papahana transcriptome holoʻokoʻa ma nā kahua noiʻi like ʻole, lawe kā mākou hui i ka waiwai o ka ʻike i kēlā me kēia papahana.
●Hoʻomalu maikaʻi loa: hoʻokō mākou i nā wahi mana koʻikoʻi ma nā pae āpau, mai ka laʻana a me ka hoʻomākaukau ʻana i ka waihona a hiki i ka sequencing a me nā bioinformatics.Mālama kēia mākaʻikaʻi ʻana i ka hāʻawi ʻana i nā hopena kiʻekiʻe mau.
● Hōʻike piha: hoʻohana mākou i nā ʻikepili he nui e hoʻopuka pono i nā Genes Differentially Expressed (DEGs) a hana i ka loiloi hoʻonui e pili ana, e hāʻawi ana i nā ʻike e pili ana i nā kaʻina kelepona a me nā molekala e pili ana i ka pane transcriptome.
●Kākoʻo ma hope o ke kūʻai aku: ʻO kā mākou kūpaʻa e ʻoi aku ma mua o ka hoʻopau ʻana o ka papahana me kahi manawa lawelawe 3 mahina ma hope o ke kūʻai aku.I kēia manawa, hāʻawi mākou i ka hahai ʻana o ka papahana, kōkua hoʻoponopono, a me nā hālāwai Q&A e hoʻoponopono i nā nīnau pili i nā hopena.
Nā Koina Laʻana a me ka Hoʻouna ʻana
| Hale Waihona Puke | Hoʻolālā kaʻina | Manaʻo ʻia ka ʻikepili | Ka Mana Mana |
| Ua pau ka rRNA | Illumina PE150 | 16 Gb | Q30≥85% |
| Nui i koho ʻia | Illumina SE50 | Heluhelu 10-20M |
Nā Koina Laʻana:
Nucleotides:
| Conc.(ng/μl) | Ka nui (μg) | Maemae | Pono pono |
| ≥ 100 | ≥ 1 | OD260/280=1.7-2.5 OD260/230=0.5-2.5 Hoʻopaʻa ʻia a ʻaʻole hoʻi i ka protein a i ʻole DNA contamination i hōʻike ʻia ma ka gel. | Nā mea kanu: RIN≥6.5 Holoholona: RIN≥7.0 5.0≥28S/18S≥1.0; palena a ʻaʻole kiʻekiʻe kumu |
Manaʻo ʻia ka hāʻawi laʻana
ipu:
2 ml centrifuge paipu (ʻaʻole ʻōlelo ʻia ʻo Tin foil)
Laʻana lepili: Pūʻulu+hoʻopiʻi e laʻa me A1, A2, A3;B1, B2, B3... ...
Hoʻouna:
1.Dry-ice: Pono e hoʻopiliʻia nā laʻana i loko o nāʻeke a kanuʻia i loko o ka hau maloʻo.
2.RNAstable tubes: Hiki ke hoʻomaloʻo ʻia nā laʻana RNA ma ka RNA stabilization tube (eg RNAstable®) a hoʻouna ʻia i loko o ka lumi wela.
Holo Hana Hana
Hoʻolālā hoʻokolohua
Hāʻawi laʻana
RNA unuhi
Kūkulu hale waihona puke
Ke kaʻina ʻana
ʻIkepili ʻikepili
Nā lawelawe ma hope o ke kūʻai aku
Bioinformatics
Hōʻike hōʻike RNA
Nā genes Hōʻike ʻokoʻa
ka nānā ʻana o ka ceRNA
Nā miRNA i hōʻike ʻokoʻa a me nā RNA pili
E ʻimi i nā holomua noiʻi i hoʻomaʻamaʻa ʻia e BMKGene 'nā lawelawe hoʻokaʻina transcriptome holoʻokoʻa ma o ka hōʻiliʻili ʻana o nā puke.
Dai, Y. et al.(2022) 'Nā palapala hōʻike piha o nā mRNA, lncRNA a me nā miRNA i ka maʻi Kashin-Beck i ʻike ʻia e ka RNA-sequencing', Molecular Omics, 18(2), pp. 154–166.doi: 10.1039/D1MO00370D.
Liu, N. nan et al.(2022) 'Ka nānā 'ana i nā transcriptomes holo'oko'a no ke kū anuanu o Apis cerana ma Changbai Mountain i ka wā ho'oilo.', Gene, 830, pp. 146503–146503.doi: 10.1016/J.GENE.2022.146503.
Wang, XJ et al.(2022) 'Multi-Omics Integration-Based Prioritization of Cometing Endogenous 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) 'Hōʻike ka ʻimi hoʻohui ʻana o ka lncRNA/circRNA-miRNA-mRNA hōʻike ʻike i nā ʻike hou i nā hana hiki ke pane i nā nematodes root-knot i ka pīkī', BMC Genomics, 23(1), pp. 1–12.doi: 10.1186 / S12864-022-08470-3 / NUI / 7.
Yan, Z. et al.(2022) 'Hōʻike ka hoʻokaʻina RNA holoʻokoʻa i nā hana molekala e pili ana i ka mālama ʻana i ka maikaʻi postharvest ma broccoli e ka ʻulaʻula LED irradiation', Postharvest Biology and Technology, 188, p.111878. doi: 10.1016/J.POSTHARVBIO.2022.111878.
