Tag Nrho-Ntev MRAND EXTRECTION -PACBio
Nta
Rdthes synthesis los ntawm poly-ib MRNA ua raws li kev npaj qiv ntawv
● Kev sib txuas hauv CCS hom, tsim cov HiFi nyeem
● Kev nthuav dav ntawm cov ntawv sau tag nrho
Kev tsom xam tsis tas yuav tsum siv cov ntsuas genome; Txawm li cas los xij, nws yuav ua haujlwm
● Bioinformatic Tsom xam thiaj li tsom xam cov ntawv teev tseg isoform lncrd, noob fenuss, poly-adenylation, thiab cov qauv noob
Kev Pabcuam Zoo
●Siab Tshaj Tawm: HiFi nyeem nrog qhov raug> 99.9% (Q30), piv rau NGS
Lwm Txoj Kev Tshawb Fawb Spicing Splicing: Kev sib txuas ntawm tag nrho cov ntawv loj pab ua kom cov lej ua tau zoo thiab cov cim
●Kev txawj ntse dav: Nrog cov ntaub ntawv teev tseg ntawm kev ua tiav ntau dua 1100 pacbio cov phiaj xwm puv ntoob thiab ua cov qauv ntev tshaj plaws thiab ua peb pab neeg coj kev muaj txiaj ntsig rau txhua qhov project.
●Kev them nyiaj yug tom qab muag: Peb cov kev cog lus txuas ntxiv tshaj qhov ua tiav nrog 3-hli tom qab muag lub sijhawm pabcuam. During this time, we offer project follow-up, troubleshooting assistance, and Q&A sessions to address any queries related to the results.
Qauv yuav tsum tau ua thiab xa khoom
| Lub tsev qiv ntawv | Seitencing Lub Tswv Yim | Cov ntaub ntawv pom zoo | Kev Tswj Xyuas Zoo |
| Polya enriched mRNS lub tsev qiv ntawv | Pacbio sequel II Pacbio Revio | 20/40 GB 5/10 m ccs | Q30-85% |
Qauv Yuav Tsum Tau Ua:
Nucleotides:
● ● Nroj tsuag:
Hauv paus, kav lossis Petal: 450 mg
Nplooj lossis noob: 300 mg
Txiv Hmab Txiv Ntoo: 1.2 g
Tsiaj:
Lub plawv lossis txoj hnyuv: 300 mg
Viscera lossis Hlwb: 240 mg
Cov leeg: 450 mg
Cov pob txha, plaub hau lossis tawv nqaij: 1g
Arthropods:
Kab: 6g
CRUSTEAA: 300 MG
Lub ntshav tag nrho: 1 raj
● Cells: 106 ig
| Conc. (NG / μl) | Tus nqi (μg) | Kev coj dawb huv | Kev muaj siab dawb paug |
| ≥ 100 | ≥ 1.0 | OD260 / 280 = 1.7-2.5 OD260 / 230 = 0.5-25-2.5 Txwv tsis pub lossis tsis muaj protein los yog cov khoom tsis huv uas muaj nyob ntawm gel. | Rau cov nroj tsuag: rin≥7.5; Rau cov tsiaj: rin≥8.0; 5.0≥ "/ 18s≥1.0; Txwv tsis pub lossis tsis muaj qhov nce hauv qab |
Pom zoo tus qauv xa mus
Thawv: 2 ml Centrifuge raj (tin ntawv ci tsis pom zoo)
Qauv kos npe: Pab pawg + hloov kho dua li AG1, A2, A3; B1, B2, B3.
Cov zaub mov xa tuaj:
1. Cov dej khov txias: Cov qauv yuav tsum tau ntim rau hauv cov hnab thiab faus rau hauv cov dej khov qhuav.
2. Rnastable Hlis: RNA cov qauv tuaj yeem qhuav hauv RNA stabilization raj (piv txwv li rnastable®) thiab xa tawm hauv chav tsev kub.
Kev Pabcuam Khiav
Kev Tshawb Fawb Kev Tshawb Fawb
Qauv Xa
RNA rho tawm
Kev Siv Hauv Tsev Qiv Ntawv
Ze
Cov Ntaub Ntawv Kev Ntsuam Xyuas
Cov Kev Pab Tom Qab Muag
Xws li kev tsom xam hauv qab no:
● Cov ntaub ntawv nyoos Zoo tswj
Lwm Txoj Kev Tshawb Fawb Polyadenyfation Kev Tshawb Xyuas (APA)
Fullion Transscript tsom xam
Lwm Txoj Kev Tshawb Fawb Spicing Splicing
● Txheeb xyuas cov thoob ntiaj teb ib daim ntawv sau orthocs (busco) tsom xam
● Cov ntawv twv kev tshawb xyuas me me Kev Tshawb Xyuas: Kev twv ua ntej ntawm coding ua ntu zus (CDS) thiab kev tshaj xo
● Lncra Kev Ntsuam Xyuas: Kev twv ua ntej ntawm LnCrna thiab Hom Phiaj
● Microatelite tus kheej (SSR)
Kev Tshawb Xyuas Kev Tshawb Xyuas Kev Loj Hlob
Lwm Txoj Kev Ntsuam Xyuas Splicing
Lwm Txoj Polyadenyylation Kev Tshawb Xyuas (APA)
Ua haujlwm tshaj tawm ntawm cov ntawv sau npe tshiab
Tshawb cov kev nce qib uas yooj yim los ntawm BMKGene's Nanopore puv-ntev mRNA cov kev pabcuam hauv cov kev pabcuam tshaj tawm no.
Ma, Y. li al. (2023) 'Kev Tshawb Xyuas Pacbio thiab ONT RNA EXTRESS TXOJ CAI RAU NEMOPILEMA NOMURAI VENOM SCHENCATION', GENOMICS, 115 (6), p. 110709. DOI: 10.1016 / J.YGARO.2023.110709.
Chao, Q. li al. (2019) 'Qhov kev loj hlob ntawm cov neeg sib txuas ntawm cov neeg Populus STEM transcriptome', cog biotechnology Journ, 17 (1), pp. 206-219. DOI: 10.1111 / PBI.12958.
Deng, H. li al. (2022) 'kev hloov pauv ntawm cov ntsiab lus ascorbic acid thiab riporbate txiv hmab txiv ntoo "thiab cov phau ntawv xov xwm thoob ntiaj teb ntawm cov molecular sciular, 23 (10), p. 5808. DOI: 10.3390 / iJMS23105808 / S1.
Hua, X. li al. (2022) Kev twv ua ntej ntawm Biosyncthetic pathway cov noob uas muaj kev cuam tshuam nrog Biophathlins ', kev sib txuas lus biology 2022 5: 1, 5 (1), pp. 1-10. DOI: 10.18 / s42003-022-03000-z.
Liu, M. li al. (2023) '' Suaq iso-seq thiab ethrice RNA-SEQ tsom xam ntawm Tuta Conccrome (MeYROME P450 Genes, 14 (4), p. 363. DOI: 10.3390 / Kab14040363 / S1.
Wang, Lijun li al. (2019) 'Kev sojntsuam ntawm transcriptome nyuaj siv pacbio ib leeg-molecule rict-lub sij hawm tshawb xyuas ntawm Ricinus Accial, BMC Genomics, 20 (1), pp. 1-17. DOI: 10.1186 / s12864-019-5832-9.
