Ulandelelwano lwe-Prokaryotic RNA
Iimbonakalo
● Ukulungiswa kwesampuli ye-RNA kwakubandakanya ukuncipha kwe-rRNA kulandele ukulungiswa kwethala leencwadi le-RNA eliya kwicala elithile.
● Uhlalutyo lwe-Bioinformatic olusekelwe ekuhambelaneni ne-genome yesalathiso
● Uhlalutyo lubandakanya ukubonakaliswa kwezakhi zofuzo kunye nee-DEG kodwa kunye nesakhiwo se-transcript kunye nohlalutyo lwe-sRNA
Iingenelo zeNkonzo
●Ulawulo Oluqinileyo Lomgangatho: siphumeza amanqaku olawulo oluphambili kuzo zonke izigaba, ukusuka ekulungiseleleni isampuli kunye nethala leencwadi ukuya kulandelelwano kunye ne-bioinformatics. Olu hlolo olucokisekileyo luqinisekisa ukunikezelwa kweziphumo ezisemgangathweni ophezulu rhoqo.
●Idatha yoLungelelaniso oluQhelekileyo: ngenxa yokuba ukulungiswa kwethala leencwadi le-RNA kubhekiselele kwicala elithile, okuvumela ukuchongwa kweengxelo ezichasene nengqiqo.
●Uhlalutyo olupheleleyo olulungiselelwe iiTranscriptomes zeProkaryotic: umbhobho we-bioinformatic awubandakanyi nje kuphela uhlalutyo lokubonakaliswa kwezakhi zofuzo kodwa nohlalutyo lwesakhiwo se-transcript, kubandakanya ukuchongwa kwee-operons, ii-UTR kunye nee-promoters. Ikwabandakanya uhlalutyo lwee-sRNA, oko kukuthi i-annotation kunye nokuqikelela isakhiwo sesibini kunye neethagethi.
●Inkxaso Emva koThengiso: ukuzibophelela kwethu kudlulela ngaphaya kokugqitywa kweprojekthi kunye nexesha lenkonzo emva kokuthengiswa kweenyanga ezi-3. Ngeli xesha, sinikezela ngokulandela iprojekthi, uncedo lokusombulula iingxaki, kunye neeseshoni ze-Q&A ukuphendula nayiphi na imibuzo enxulumene neziphumo.
Iimfuno zeSampuli kunye nokuHanjiswa
| Ithala leeNcwadi | Icebo lokulandelelanisa izinto | Idatha iyacetyiswa | Ulawulo lwemeko |
| ilayibrari yesikhokelo ephelileyo ye-rRNA | I-Illumina PE150 | 1-2 Gb | Q30≥85% |
Iimfuno zeSampuli:
| I-Conc.(ng/μl) | Ubungakanani (μg) | Ubunyulu | Ingqibelelo |
| ≥ 50 | ≥ 1 | OD260/280=1.8-2.0 OD260/230=1.0-2.5 Ungcoliseko lweproteni okanye lwe-DNA oluncinci okanye alubonakali kwijeli. | I-RIN≥6.5 |
Ukuhanjiswa kweSampuli okuCetyisiweyo
Isikhongozeli: Ityhubhu ye-centrifuge ye-2 ml (Akukhuthazwa ukusebenzisa i-tin foil)
Iilebheli zesampulu: Iqela + ikopi umz. A1, A2, A3; B1, B2, B3.
Ukuthunyelwa:
1. Umkhenkce owomileyo: Iisampulu kufuneka zipakishwe kwiingxowa zize zingcwatywe kumkhenkce owomileyo.
2. Iityhubhu ze-RNAstable: Iisampulu ze-RNA zingomiswa kwityhubhu yokuzinzisa i-RNA (umz. i-RNAstable®) kwaye zithunyelwe kubushushu begumbi.
Ukuhamba komsebenzi wenkonzo
Ukuhanjiswa kwesampulu
Ukwakhiwa kweThala leeNcwadi
Ukulandelelana
Uhlalutyo lwedatha
Iinkonzo zasemva kokuthengisa
Ukuhamba komsebenzi wohlalutyo lwe-Bioinformatic
Kubandakanya uHlahlelo Olulandelayo:
● Ulawulo lomgangatho wedatha eluhlaza
● Ukulungelelaniswa kwi-genome yesalathiso
● Uvavanyo loMgangatho weThala leeNcwadi: Ukuqhekeka kwe-RNA ngokungacwangciswanga, ubungakanani bokufaka kunye nokulandelelana kokugcwalisa
● Inkcazo esebenzayo yezakhi zofuzo eziqikelelweyo zokubhala ikhowudi
● Uhlalutyo lweenkcazo: ulwalamano kunye noHlalutyo lweeCandelo eziPhambili (i-PCA)
● Ukubonakaliswa kweGene eyahlukileyo (ii-DEG)
● Inkcazo esebenzayo kunye nokuphucula ii-DEG
● Uhlalutyo lwe-sRNA: ukuqikelela, isichazi-magama, injongo, kunye nokuqikelela ulwakhiwo lwesibini
● Uhlalutyo lwesakhiwo seTranscript: ii-operon, izikhundla zokuqala nezokugqibela, ummandla ongaguqulelwanga (UTS), umkhuthazi, kunye nohlalutyo lwe-SNP/InDel
Ukulandelelanisa ukugcwala
Isichazi-magama esisebenzayo seejini zokubhala ikhowudi
Ulwalamano phakathi kweesampuli
Uhlalutyo lweeGene eziVezwayo ezahlukeneyo (ii-DEG)
Uhlalutyo lokuphucula ukusebenza
isichazi-magama se-sRNA
Hlola uphuhliso oluququzelelwe ziinkonzo ze-BMKGene's Nanopore full-length mRNA sequencing kule ncwadi iphambili.
UGuan, CP et al. (2018) 'Utshintsho lweTranscriptome lweHlabathi Jikelele lwe-Biofilm-Forming Staphylococcus epidermidis Ephendula kwi-Total Alkaloids ye-Sophorea alopecuroides',Ijenali yasePoland yeMicrobiology, 67(2), iphe. 223. doi: 10.21307/PJM-2018-024.
