Kev Txheeb Xyuas Prokaryotic RNA
Cov yam ntxwv
● Kev ua cov qauv RNA suav nrog kev tshem tawm rRNA ua ntej thiab tom qab ntawd npaj cov tsev qiv ntawv RNA.
● Kev tshuaj xyuas bioinformatic raws li kev sib phim rau cov genome siv
● Kev tshuaj xyuas suav nrog kev qhia tawm cov noob caj noob ces thiab DEGs tab sis kuj tseem muaj cov qauv sau ntawv thiab kev tshuaj xyuas sRNA
Cov Kev Pabcuam Zoo
●Kev Tswj Xyuas Zoopeb siv cov ntsiab lus tswj hwm tseem ceeb thoob plaws txhua theem, txij li kev npaj cov qauv thiab tsev qiv ntawv mus rau kev txheeb xyuas thiab bioinformatics. Qhov kev saib xyuas zoo no ua kom ntseeg tau tias muaj cov txiaj ntsig zoo tas li.
●Cov Ntaub Ntawv Sequencing Tshwj Xeeb ntawm Strand: vim yog RNA lub tsev qiv ntawv npaj ua qhov kev taw qhia, ua rau muaj kev txheeb xyuas cov ntawv sau tsis zoo.
●Kev Tshawb Fawb Tag Nrho Uas Tsim Los Rau Prokaryotic Transcriptomes: cov raj xa dej bioinformatic tsis yog tsuas yog suav nrog kev tshuaj xyuas ntawm cov noob caj noob ces xwb tab sis kuj tseem muaj kev tshuaj xyuas cov qauv sau ntawv, suav nrog kev txheeb xyuas cov operons, UTRs thiab cov neeg txhawb nqa. Nws kuj suav nrog kev tshuaj xyuas ntawm sRNAs, uas yog kev sau ntawv thiab kev kwv yees ntawm cov qauv thib ob thiab cov hom phiaj.
●Kev Txhawb Nqa Tom Qab Muag Khoom: peb txoj kev cog lus txuas ntxiv mus dhau qhov ua tiav qhov project nrog rau 3 lub hlis tom qab muag kev pabcuam. Thaum lub sijhawm no, peb muab kev saib xyuas 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 depleted directional library | Illumina PE150 | 1-2 Gb | Q30≥85% |
Cov Qauv Yuav Tsum Tau Ua:
| Conc.(ng/μl) | Tus nqi (μg) | Kev dawb huv | Kev ncaj ncees |
| ≥ 50 | ≥ 1 | OD260/280=1.8-2.0 OD260/230=1.0-2.5 Muaj tsawg los yog tsis muaj protein lossis DNA uas pom ntawm gel. | RIN≥6.5 |
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 tuaj rau hauv chav tsev kub.
Kev Ua Haujlwm Pabcuam
Kev xa khoom qauv
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 Ua Haujlwm Tshawb Fawb Txog Bioinformatic
Muaj xws li Kev Tshawb Fawb Hauv Qab No:
● Kev tswj xyuas cov ntaub ntawv raw zoo
● Kev sib phim rau cov genome siv los piv txwv
● Kev Ntsuam Xyuas Qhov Zoo ntawm Tsev Qiv Ntawv: RNA fragmentation randomness, qhov loj ntawm cov ntxig thiab sequencing saturation
● Kev piav qhia txog kev ua haujlwm ntawm cov noob caj noob ces uas tau kwv yees ua ntej
● Kev Tshawb Fawb Txog Kev Qhia: kev sib raug zoo thiab Kev Tshawb Fawb Txog Cheebtsam Tseem Ceeb (PCA)
● Kev Qhia Txog Cov Gene Txawv (DEGs)
● Kev piav qhia txog kev ua haujlwm thiab kev txhim kho ntawm DEGs
● Kev Tshuaj Xyuas sRNA: kev kwv yees, kev piav qhia, lub hom phiaj, thiab kev kwv yees qauv theem ob
● Kev Tshuaj Xyuas Cov Qauv ntawm Cov Ntawv Sau: operons, qhov chaw pib thiab xaus, Thaj chaw tsis tau txhais (UTS), tus txhawb nqa, thiab kev tshuaj xyuas SNP/InDel
Kev ntsuas qhov saturation
Kev piav qhia txog kev ua haujlwm ntawm cov noob caj noob ces coding
Kev sib raug zoo ntawm cov qauv
Kev tshuaj xyuas cov noob caj noob ces sib txawv (DEGs)
Kev tshuaj xyuas kev nplua nuj ntawm kev ua haujlwm
cov lus piav qhia ntawm sRNA
Tshawb nrhiav cov kev nce qib uas tau pab txhawb los ntawm BMKGene's Nanopore full-length mRNA sequencing cov kev pabcuam hauv phau ntawv tshaj tawm tshwj xeeb no.
Guan, CP et al. (2018) 'Kev Hloov Pauv Thoob Ntiaj Teb ntawm Biofilm-Forming Staphylococcus epidermidis Teb rau Tag Nrho Alkaloids ntawm Sophorea alopecuroides',Phau Ntawv Xov Xwm Polish ntawm Microbiology, 67(2), p. 223. doi: 10.21307/PJM-2018-024.
