Liʻiliʻi RNA sequencing-Illumina
Nā hiʻohiʻona
● Koho nui o RNA ma mua o ka hoʻomākaukau ʻana i ka waihona
● ʻIkepili bioinformatic e pili ana i ka wānana miRNA a me kā lākou mau pahuhopu
Pono lawelawe
●ʻIkepili bioinformatics piha:hiki i ka ʻike ʻana i nā miRNA i ʻike ʻia a me ka moʻolelo, ka ʻike ʻana i nā pahuhopu miRNAs a me nā annotation hana kūpono a me ka hoʻonui ʻana me nā ʻikepili lehulehu (KEGG, GO)
●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.
●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.
●ʻIke Nui: me kahi moʻolelo o ka hoʻopaʻa kūleʻa ʻana ma luna o nā papahana sRNA he nui e uhi ana ma luna o 100 mau ʻano i loko o nā kikowaena noiʻi like ʻole, lawe kā mākou hui i ka waiwai o ka ʻike i kēlā me kēia papahana.
Nā Koina Laʻana a me ka Hoʻouna ʻana
| Hale Waihona Puke | Papahana | Manaʻo ʻikepili | ʻIkepili QC |
| Nui i koho ʻia | Illumina SE50 | Heluhelu 10M-20M | Q30≥85% |
Nā Koina Laʻana:
Nucleotides:
| Conc.(ng/μl) | Ka nui (μg) | Maemae | Pono pono |
| ≥ 80 | ≥ 0.5 | 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. | RIN≥6.5; 5.0≥28S/18S≥1.0; palena a ʻaʻole kiʻekiʻe kumu |
● Nā mea kanu:
Aʻa, ʻaʻa a i ʻole Petal: 450 mg
Lau a Hua paha: 300 mg
Hua: 1.2 g
● holoholona:
Naʻau a i ʻole ka ʻōpū: 450 mg
Viscera a i ʻole lolo: 240 mg
ʻiʻo: 600 mg
Iwi, lauoho a ʻili paha: 1.5g
● Nā ʻāʻī:
Nā Insect: 9g
Crustacea: 450 mg
● Ke koko holoʻokoʻa: 2 paipu
● Nā Pūnaewele: 106 pūnaewele
● Serum a me ka plasma:6 mL
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
Ka ʻike ʻana o ka miRNA: ke ʻano a me ka hohonu
Hōʻike ʻokoʻa o ka miRNA - hui pū ʻana hiearchical
ʻO ka hōʻike kikoʻī o ka pahuhopu o nā miRNA i hōʻike ʻokoʻa
E ʻimi i nā holomua noiʻi i hoʻomaʻamaʻa ʻia e BMKGene 'sRNA sequencing services ma o ka hōʻiliʻili curated o nā puke.
Chen, H. et al.(2023) 'O nā ma'i ma'i ma'i ke'ake'a i ka biosynthesis saponin a me ka photosynthesis ma Panax notoginseng', Plant Physiology and Biochemistry, 203, p.108038. doi: 10.1016/J.PLAPHY.2023.108038.
Li, H. et al.(2023) 'O ka mea kanu FYVE domain-i loko o ka protein FREE1 hui pū me nā mea microprocessor e kāohi i ka miRNA biogenesis', hōʻike EMBO, 24(1).doi: 10.15252/EMBR.202255037/SUPPL_FILE/EMBR202255037-SUP-0004-SDATAFIG4.TIF.
Yu, J. et al.(2023) 'Hoʻomalu ka MicroRNA Ame-Bantam-3p i ka hoʻomohala ʻana o ka pupal larval ma o ka hoʻopaʻa ʻana i ka nui o ka Epidermal Growth Factor-like Domains 8 Gene (megf8) i ka Honeybee, Apis mellifera', International Journal of Molecular Sciences, 24(6), p .5726. doi: 10.3390/IJMS24065726/S1.
Zhang, M. et al.(2018) 'Integrated Analysis of MiRNA and Genes Associated with Meat Quality Reveals that Gga-MiR-140-5p Affects Intramuscular Fat Deposition in Chickens', Cellular Physiology and Biochemistry, 46(6), pp. 2421–2433.doi: 10.1159/000489649.
