Ngena kwi-BMKCloud

Iimveliso

Ukulandelelana kwe-RNA encinci ye-Exosomal-Illumina

Umfanekiso oPhambili we-Exosomal Small RNA Sequencing-Illumina

Ii-Exosomes zii-vesicles ezincinci ezikhutshwa ziiseli, zihlala zinobubanzi obuphakathi kwee-nanometers ezingama-30 ukuya kwi-100. Ezi vesicles ziqulathe ii-RNA ezahlukeneyo. Phakathi kweentlobo ze-RNA kwii-exosomes, eyona ixhaphakileyo nefundwa kakhulu yi-microRNA (miRNA). I-miRNA luhlobo lwee-RNA ezincinci ezingabhalisiyo iikhowudi ezimalunga ne-18-25 nucleotides ubude. Zilawula ukuthuliswa kwe-gene emva kokubhalwa ngokubophelela kwindawo engama-3′ engaguqulelwanga (3′ UTR) yee-mRNA ezijoliswe kuzo, ngaloo ndlela zilawula ukubonakaliswa kwe-gene. Umzekelo, ii-exosomes ezikhutshwa ziiseli ezithile ze-tumor ziqulathe ii-miRNA ezithile, ezifana ne-miR-126 kunye ne-miR-92a. Ezi miRNAs zinokuchaphazela ukubonakaliswa kwe-gene kwiiseli ezamkelayo kwaye zikhuthaze i-tumor angiogenesis (Tomohiro Umezu, et al., Oncogene, 2012).

Qhagamshelana nathi

Iinkcukacha zeNkonzo

i-bioinformatics

Iziphumo zeDemo

Iimbonakalo

● I-RNA iyonke ephuma e-exosomal isetyenziswa njengezinto zokufaka. Ngexesha lokuhlanjululwa, ii-miRNA Beads zisetyenziswa ukwahlula i-miRNA.

● Uhlalutyo lwe-Bioinformatic lujolise ekuqikeleleni i-miRNA kunye neenjongo zayo.

Iingenelo zeNkonzo

● Uhlalutyo olupheleleyo lwe-bioinformatics: Ukuvumela ukuchongwa kwee-miRNA ezaziwayo nezintsha, ukuchongwa kweenjongo ze-miRNA, kunye nokuchazwa okusebenzayo kunye nokuphuculwa ngeedathabheyisi ezininzi (KEGG, GO).

● Ulawulo Oluqinileyo Lomgangatho: Sisebenzisa 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.

● Inkxaso Emva Kokuthengisa: Ukuzinikela kwethu kudlulela ngaphaya kokugqitywa kweprojekthi kunye nexesha leenkonzo emva kokuthengisa leenyanga 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	Iqonga	Idatha ecetyiswayo	Idatha ye-QC
Ukwahlulwa kwe-miRNA engaphandle	I-Illumina SE50	Ukufundwa kwe-10M-20M	Q30≥85%

Iimfuno zeSampuli

Iinukleotide:

Isixa (ng)	Umthamo (μL)
40	20

Izicubu:

I-Exosome: 100 μL; 40 μg; >10^9 amasuntswana/mL
I-serum/plasma: 5 mL
I-supernatant yenkcubeko yeseli: 100 mL
Umchamo: 100 mL
Amathe: 15 mL
Ukukhupha iimpumlo: 15 mL
Ulwelo lweCerebrospinal: 15 mL
Ulwelo lwe-Amniotic: 50 mL
Amadlozi: 5 mL
Inyongo: 5 mL
Ii-Ascites: 50 mL
Ubisi: 15 mL
Ulwelo lweFollicular: 15 mL
I-supernatant yelindle: 50 mL
I-supernatant yenkcubeko ye-microbial: 100 mL
Izicubu zezilwanyana: 3 g
Izicubu zesityalo: 1 g

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

Uyilo lovavanyo

Ukuhanjiswa kwesampulu

Ukukhupha i-RNA

Ukwakhiwa kweThala leeNcwadi

Ukulandelelana

Uhlalutyo lwedatha

Iinkonzo zasemva kokuthengisa

Ngaphambili: I-BMKMANU S3000_Spatial Transcriptome

Okulandelayo: I-Exosomal mRNA/LncRNA/CirrcRNA Sequencing-Illumina

I-Bioinformatics

● Ulawulo lomgangatho wedatha eluhlaza
● udidi lwe-sRNA
● Ukulungelelaniswa kwi-genome ebhekisa kwi-genome
● Ukuchongwa kwe-miRNA eyaziwayo nentsha
● Uhlalutyo lokubonakaliswa kwe-miRNA eyahlukileyo
● Inkcazo esebenzayo yeethagethi ze-miRNA

Ukuchongwa kwe-miRNA: isakhiwo kunye nobunzulu