Kev Txheeb Xyuas Tag Nrho Cov Ntawv Sau - Illumina
Cov yam ntxwv
● Kev tsim ob lub tsev qiv ntawv los ua kom tiav cov ntawv sau ua ke: tsev qiv ntawv rRNA depletion thiab kev npaj tsev qiv ntawv RNA me me.
● Kev tshuaj xyuas bioinformatic tiav ntawm mRNA, lncRNA, circRNA thiab miRNA hauv cov ntawv qhia bioinformatic sib cais, ntxiv rau kev tshuaj xyuas ua ke nrog txhua qhov kev qhia tawm RNA ua ke, suav nrog kev tshuaj xyuas ceRNA networks.
Cov Kev Pabcuam Zoo
●Kev Paub Ntau YamPeb tau ua tiav ntau tshaj 2100 qhov project transcriptome, uas suav nrog ntau hom qauv sib txawv. Peb coj kev txawj ntse ntau yam los rau txhua qhov project.
●Kev Tswj Xyuas ZooPeb siv cov ntsiab lus tswj hwm tseem ceeb thoob plaws txhua theem, txij li kev npaj cov qauv mus rau kev npaj tsev qiv ntawv, kev txheeb xyuas cov ntaub ntawv thiab kev siv bioinformatics. Peb qhov kev saib xyuas zoo ua kom ntseeg tau tias muaj cov txiaj ntsig zoo tas li.
●Cov Lus Qhia Txog Kev Ua HaujlwmPeb siv ntau lub database los sau cov Differentially Expressed Genes (DEGs) thiab ua cov kev tshuaj xyuas kev nplua nuj. Txoj hauv kev no muab kev nkag siab txog cov txheej txheem cellular thiab molecular uas yog lub hauv paus ntawm cov lus teb transcriptome, kom ntseeg tau tias koj tau txais txhua cov ntaub ntawv hais txog koj cov ntaub ntawv sim.
●Kev Tshawb Fawb Txog Kev Tswj Xyuas Hauv Is Taws NemKev tshuaj xyuas lub network ceRNA yog ua tau los ntawm kev sib koom ua ke ntawm mRNA, lncRNA, circRNA, thiab miRNA thiab los ntawm kev ua haujlwm bioinformatic tag nrho.
●Kev Txhawb Nqa Tom Qab Muag KhoomPeb nkag siab tias qhov tseem ceeb ntawm kev nyob ntawd yog dab tsi, yog vim li cas peb txoj kev cog lus txuas ntxiv mus dhau qhov ua tiav qhov project nrog 3 lub hlis kev pabcuam tom qab muag. Thaum lub sijhawm no, peb muab kev saib xyuas qhov project, kev pab daws teeb meem, thiab cov lus nug thiab lus teb los teb cov lus nug ntsig txog cov txiaj ntsig.
Cov Qauv Yuav Tsum Tau Ua thiab Kev Xa Khoom
| Tsev qiv ntawv | Txoj kev npaj ua sequencing | Cov ntaub ntawv pom zoo | Kev Tswj Xyuas Zoo |
| rRNA ploj mus | Illumina PE150 | 16 Gb | Q30≥85% |
| Qhov loj uas tau xaiv | Illumina SE50 | 10-20M nyeem |
Cov Qauv Yuav Tsum Tau Ua:
Cov nucleotides:
| Conc.(ng/μl) | Tus nqi (μg) | Kev dawb huv | Kev ncaj ncees |
| ≥ 80 | ≥ 1.6 | OD260/280=1.7-2.5 OD260/230=0.5-2.5 Muaj tsawg los yog tsis muaj protein lossis DNA uas pom ntawm gel. | RIN≥6.0 5.0≥28S / 18S≥1.0; txwv los yog tsis muaj qhov siab pib |
Kev Xa Qauv Pom Zoo
Lub thawv: 2 ml centrifuge tube (Tsis pom zoo kom siv ntawv ci)
Piv txwv kev sau npe: Pawg + theej piv txwv li A1, A2, A3; B1, B2, B3.
Kev Xa Khoom:
1. Dej khov qhuav: Cov qauv yuav tsum tau ntim rau hauv hnab thiab faus rau hauv dej khov qhuav.
2. Cov raj RNAstable: Cov qauv RNA tuaj yeem ziab hauv cov raj RNA stabilization (piv txwv li RNAstable®) thiab xa mus rau qhov kub thiab txias hauv chav.
Kev Ua Haujlwm Pabcuam
Tsim qauv kev sim
Kev xa khoom qauv
Kev rho tawm RNA
Kev tsim kho tsev qiv ntawv
Kev txheeb xyuas kab ke
Kev tshuaj xyuas cov ntaub ntawv
Cov kev pab cuam tom qab muag
Kev tshawb fawb txog bioinformatics
Ntxiv rau txhua qhov kev tshuaj xyuas ntawm txhua qhov sib txawv, 4 hom RNA tau muab tso ua ke rau hauv peb qhov kev tshuaj xyuas ua ke:
- Kev tshuaj xyuas kev sib koom ua ke ntawm txhua hom ntaub ntawv RNA sib txawv
- Kev piav qhia txog kev qhia tawm RNA
- Kev piav qhia txog kev qhia sib txawv
- CeRNA Network
- Cov noob caj noob ces tseem ceeb uas tau koom ua ke hauv txoj kev
- Kev tshuaj xyuas kev txuag
- Kev tsom mus rau kev sib raug zoo
Kev piav qhia txog kev qhia tawm RNA
Cov noob caj noob ces sib txawv
kev tshuaj xyuas ceRNA
Cov miRNAs uas qhia tawm sib txawv thiab cov RNAs cuam tshuam
Tshawb nrhiav kev nce qib ntawm kev tshawb fawb uas tau pab txhawb los ntawm BMKGene 'tag nrho cov kev pabcuam transcriptome sequencing los ntawm kev sau cov ntawv tshaj tawm.
Dai, Y. et al. (2022) 'Cov qauv qhia txog kev qhia txog mRNAs, lncRNAs thiab miRNAs hauv kab mob Kashin-Beck uas tau txheeb xyuas los ntawm RNA-sequencing',Molecular Omics, 18(2), pp. 154–166. doi: 10.1039/D1MO00370D.
Liu, N. nan et al. (2022) 'Kev tshuaj xyuas cov ntawv sau tag nrho ntawm kev tiv taus txias ntawm Apis cerana hauv Changbai Roob thaum lub sijhawm overwintering.',Cov noob caj noob ces, 830, nplooj ntawv 146503–146503. doi: 10.1016/J.GENE.2022.146503.
Wang, XJ et al. (2022) 'Kev Sib Koom Tes Ntau-Omics-Raws Li Qhov Tseem Ceeb ntawm Kev Sib Tw Endogenous RNA Regulation Networks hauv Small Cell Lung Cancer: Cov Yam Ntxwv Molecular thiab Cov Neeg Sib Tw Tshuaj',Cov ciam teb hauv Oncology, 12, ib. 904865. doi: 10.3389/FONC.2022.904865/BIBTEX.
Xu, P. et al. (2022) 'Kev tshuaj xyuas ua ke ntawm cov qauv qhia tawm ntawm lncRNA/circRNA-miRNA-mRNA qhia txog kev nkag siab tshiab rau cov txheej txheem muaj peev xwm teb rau cov nematodes hauv cov txiv laum huab xeeb',BMC Genomics, 23(1), nplooj ntawv 1–12. doi: 10.1186/S12864-022-08470-3/FIGURES/7.
Yan, Z. et al. (2022) 'Kev txheeb xyuas tag nrho cov RNA hauv kev sau qoob loo qhia txog cov txheej txheem molecular uas cuam tshuam nrog kev tswj hwm qhov zoo tom qab sau qoob loo hauv broccoli los ntawm kev siv hluav taws xob LED liab',Kev Tshawb Fawb Txog Biology thiab Technology Tom Qab Sau Qoob Loo, 188, nplooj 111878. doi: 10.1016/J.POSTHARVBIO.2022.111878.
