Prokaryotic RNA Sequencing
Likaroloana
● RNA sampole tshebetso e ne e akarelletsa ho fokotseha ha rRNA e lateloang ke tataiso ea tataiso ea laebrari ea RNA.
● Tlhahlobo ea bioinformatic e thehiloeng ho ikamahanya le genome ea litšupiso
● Tlhahlobo e kenyelletsa liphatsa tsa lefutso le li-DEGs empa hape le sebopeho sa transcript le tlhahlobo ea sRNA
Melemo ea Tšebeletso
●Taolo ea Boleng e Tiileng: re kenya tšebetsong lintlha tsa mantlha tsa taolo methating eohle, ho tloha ho lisampole le boitokiso ba laeborari ho isa ho tatellano le bioinformatics. Tlhokomelo ena e hlokolosi e tiisa hore ho fanoa ka liphetho tsa boleng bo holimo kamehla.
●Lintlha tsa Tatelano ea Strand-specific: ka lebaka la boitokiso ba laeborari ea RNA e le tataiso, e nolofalletsang ho tsebahatsa lingoliloeng tse khahlanong le maikutlo.
●Tlhahlobo e Felletseng e Lokisitsoeng ho Li-Transcriptomes tsa Prokaryotic: pipeline ea bioinformatic ha e akarelletse feela tlhahlobo ea liphatsa tsa lefutso empa hape le tlhahlobo ea sebopeho sa transcript, ho kenyelletsa le ho tsebahatsa li-operens, li-UTR le bahlahisi. E boetse e kenyelletsa tlhahlobo ea li-sRNA, e leng tlhaloso le ponelopele ea sebopeho sa bobeli le sepheo.
●Tšehetso ea Post-Sales: Boitlamo ba rona bo fetela ka nģ'ane ho ho phethoa ha morero ka nako ea tšebeletso ea likhoeli tse 3 ka mor'a thekiso. Nakong ena, re fana ka tatellano ea projeke, thuso ea ho rarolla mathata, le linako tsa Q&A ho araba lipotso life kapa life tse amanang le sephetho.
Litlhoko tsa Mohlala le Phano
| Laebrari | Leano la tatellano | Lintlha li khothalelitsoe | Taolo ea Boleng |
| rRNA e feletsoe ke laebrari ea tataiso | Illumina PE150 | 1-2 Gb | Q30≥85% |
Litlhoko tsa Mohlala:
| Conc.(ng/μl) | Chelete (μg) | Bohloeki | Botšepehi |
| ≥50 | ≥1 | OD260/280=1.8-2.0 OD260/230=1.0-2.5 Tšoaetso e fokolang kapa ha e na protheine kapa DNA e bontšitsoeng ho gel. | RIN≥6.5 |
Tlhahiso ea Mohlala e khothalelitsoeng
Setshelo: 2 ml centrifuge tube (Tin foil ha e khothalletsoe)
Mohlala oa ho ngola: Sehlopha+katiso mohlala A1, A2, A3; B1, B2, B3.
Thomello:
1. Leqhoa le omileng: lisampole li lokela ho kenngoa ka mekotleng ebe li patoa ka leqhoa le omileng.
2. Li-tubes tsa RNAstable: lisampole tsa RNA li ka omisoa ka har'a tube ea ho tsitsisa ea RNA (mohlala, RNAstable®) le ho romelloa ka mocheso oa kamore.
Phallo ea Mosebetsi oa Ts'ebeletso
Mohlala oa thomello
Kaho ea laebrari
Tatelano
Tlhahlobo ea lintlha
Litšebeletso tsa ka mor'a thekiso
Bioinformamatic Analysis Workflow
E kenyelletsa Tlhahlobo e Latelang:
● Taolo ea boleng ba data e tala
● Ho ikamahanya le genome ea litšupiso
● Tekolo ea Boleng ba Laeborari: Ho aroloa ha RNA ka tšohanyetso, ho kenya boholo le tatellano ea tlhophiso.
● Tlhaloso ea tšebetso ea liphatsa tsa lefutso tsa khouto tse boletsoeng esale pele
● Tlhaloso ea Phatlalatso: Correlation le Principal Component Analysis (PCA)
● Differential Gene Expression (DEGs)
● Tlhaloso ea ts'ebetso le ntlafatso ea li-DEGs
● Tlhahlobo ea sRNA: ho bolela esale pele, tlhaloso, sepheo, le tlhaloso ea bobeli ea sebopeho
● Tlhahlobo ea Sebopeho sa Transcript: li-operon, maemo a qalang le a ho qetela, sebaka se sa fetoletsoeng (UTS), mothetsi, le tlhahlobo ea SNP/InDel.
Sequencing saturation
Tlhaloso e sebetsang ea liphatsa tsa lefutso tsa khouto
Kamano pakeng tsa disampole
Differential Expressed Genes (DEGs) tlhahlobo
Tlhahlobo ea ts'ebetso ea ntlafatso
Tlhaloso ea sRNA
Lekola tsoelo-pele e tsamaisoang ke BMKGene's Nanopore ea bolelele bo felletseng ba litšebeletso tsa tatellano ea mRNA khatisong ena e hlahang.
Guan, CP le ba bang. (2018) 'Liphetoho tsa Global Transcriptome tsa Biofilm-Forming Staphylococcus epidermidis Ho Arabela Kakaretso ea Alkaloids ea Sophorea alopecuroides',Polish Journal ea Microbiology, 67(2), leq. 223. doi: 10.21307/PJM-2018-024.
