circRNA sequencing-Illumina
Likaroloana
● Ho fokotseha ha rRNA ho lateloa ke ho lokisetsa laebrari ea tataiso, ho nolofalletsa data ea tatellano e khethehileng ea strand.
● Bioinformamatic workflow e thusa ho bolela esale pele ka circRNA le tlhaloso ea mantsoe
Melemo ea Tšebeletso
●Lilaeborari tsa RNA tse felletseng:re sebelisa rRNA depletion ho fapana le ho fokotseha ha RNA ka mokhoa o ts'oanang ho lokiseng laebrari ea rona pele ho laeborari, ho netefatsa hore data ea tatellano ha e akarelletse feela circRNA empa hape le mRNA le lncRNA, e nolofalletsang tlhahlobo e kopaneng ho li-datasets tsena.
●Tlhahlobo ea boikhethelo ea marang-rang a tlholisano a endogenous RNA (ceRNA).: ho fana ka lintlha tse tebileng tsa mekhoa ea tsamaiso ea cellular
●Tsebo e Pharalletseng: ka rekoto e ntle ea ho sebetsana le lisampole tse fetang 20,000 ho BMK, tse nkang mefuta e fapaneng ea sampole le merero ea lncRNA, sehlopha sa rona se tlisa boiphihlelo bo bongata morerong o mong le o mong.
●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.
● 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 | Sethala | Lintlha tse khothalelitsoeng | Lintlha tsa QC |
| Poly A e atlehile | Illumina PE150 | 16-20 Gb | Q30≥85% |
Litlhoko tsa Mohlala:
Li-nucleotides:
| Conc.(ng/μl) | Chelete (μg) | Bohloeki | Botšepehi |
| ≥ 100 | ≥ 0.5 | OD260/280=1.7-2.5 OD260/230=0.5-2.5 Tšoaetso e fokolang kapa ha e na protheine kapa DNA e bontšitsoeng ho gel. | Bakeng sa limela: RIN≥6.5; Bakeng sa liphoofolo: RIN≥7.0; 5.0≥28S/18S≥1.0; bophahamo bo fokolang kapa bo se nang motheo |
● Limela:
Motso, Stem kapa Petal: 450 mg
Lekhasi kapa Peō: 300 mg
Litholoana: 1.2 g
● Phoofolo:
Pelo kapa mala: 450 mg
Viscera kapa Boko: 240 mg
mesifa: 600 mg
Masapo, Moriri kapa Letlalo: 1.5g
● Arthropods:
Likokoanyana: 9g
Crustacea: 450 mg
● Mali a feletseng:2 li-tubes
● Lisele: 106 lisele
● Serum le Plasma6ml ka
Tlhahiso ea Mehlala 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
Moqapi oa liteko
Mohlala oa thomello
Tlhahiso ea RNA
Kaho ea laebrari
Tatelano
Tlhahlobo ea lintlha
Litšebeletso tsa ka mor'a thekiso
Bioinformatics
ponelopele ea circRNA: kabo ea chromosomal
Li-circRNA tse hlahisitsoeng ka mokhoa o fapaneng - morero oa seretse se chesang
Li-circRNA tse hlahisitsoeng ka mokhoa o fapaneng - lihlopha tsa maemo a holimo
Ntlafatso e sebetsang ea liphatsa tsa lefutso tsa circRNA
Lekola tsoelopele ea lipatlisiso e tsamaisoang ke litšebeletso tsa tatellano ea BMKGene' circRNA ka pokello e hlophisitsoeng ea lingoliloeng.
Wang, X. et al.(2021) 'CPSF4 e laola sebopeho sa circRNA le microRNA mediated gene silenceing ka hepatocellular carcinoma', Oncogene 2021 40: 25, 40 (25), maq. 4338-4351.doi: 10.1038/s41388-021-01867-6.
Xia, K. et al.(2023) 'X oo-responsive transcriptome e senola karolo ea RNA133 e chitja ho hanyetsa mafu ka ho laola polelo ea OsARAB ka raese', Phytopathology Research, 5 (1), maq. 1-14.doi: 10.1186/S42483-023-00188-8/FIGURES/6.
Y, H. et al.(2023) 'CPSF3 e fetola ho leka-lekana ha litlaleho tse chitja le tse nang le linear ka hepatocellular carcinoma'.doi: 10.21203/RS.3.RS-2418311/V1.
Zhang, Y. et al.(2023) 'Tlhahlobo e feletseng ea li-circRNAs ka lefu la pelo ea sebete pele le ka mor'a ho fetisetsa sebete', International Immunopharmacology, 114, leq.109495. doi: 10.1016/J.INTIMP.2022.109495.
