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Cov khoom

Kev Tshawb Fawb Txog RNA Me Me Exosomal-Illumina

Exosomal Small RNA Sequencing-Illumina Featured Image

Exosomes yog cov vesicles me me uas cov cell tso tawm, feem ntau muaj txoj kab uas hla ntawm 30 txog 100 nanometers. Cov vesicles no muaj ntau yam RNA. Ntawm cov hom RNA hauv exosomes, qhov feem ntau thiab kawm ntau yog microRNA (miRNA). miRNA yog ib chav kawm ntawm cov RNAs me me uas tsis yog coding thiab kwv yees li 18-25 nucleotides ntev. Lawv ua haujlwm nruab nrab ntawm kev kaw cov noob caj noob ces tom qab kev sau los ntawm kev khi rau 3′ untranslated cheeb tsam (3′ UTR) ntawm cov mRNAs uas xav tau, yog li tswj kev qhia noob caj noob ces. Piv txwv li, exosomes uas qee cov cell qog nqaij hlav tso tawm muaj cov miRNAs tshwj xeeb, xws li miR-126 thiab miR-92a. Cov miRNAs no tuaj yeem cuam tshuam kev qhia noob caj noob ces hauv cov cell txais thiab txhawb kev loj hlob ntawm qog nqaij hlav (Tomohiro Umezu, et al., Oncogene, 2012).

Tiv Tauj Peb

Cov Lus Qhia Txog Kev Pabcuam

kev kawm txog bioinformatics

Cov Txiaj Ntsig Qhia Txog Kev Ua Qauv

Cov yam ntxwv

● Exosomal tag nrho RNA yog siv ua cov khoom siv nkag. Thaum lub sijhawm ntxuav, miRNA Beads raug siv los cais miRNA.

● Kev tshuaj xyuas bioinformatic tsom mus rau kev kwv yees miRNA thiab lawv cov hom phiaj.

Cov Kev Pabcuam Zoo

● Kev tshuaj xyuas bioinformatics kom ntxaws: Pab kom paub txog ob qho tib si miRNAs uas paub thiab tshiab, kev paub txog cov hom phiaj miRNAs, thiab kev sau ntawv ua haujlwm thiab kev txhim kho nrog ntau lub ntaub ntawv (KEGG, GO).

● Kev Tswj Xyuas Zoo: Peb siv cov ntsiab lus 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.

● Kev Pab Tom Qab Muag Khoom: Peb txoj kev cog lus txuas ntxiv mus dhau qhov project ua tiav nrog 3 lub hlis kev pabcuam tom qab muag khoom. 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	Platform	Cov ntaub ntawv pom zoo	Kev Tswj Xyuas Cov Ntaub Ntawv
Kev rho tawm ntawm exosomal miRNA	Illumina SE50	10M-20M nyeem	Q30≥85%

Cov Qauv Yuav Tsum Tau Ua

Cov nucleotides:

Tus nqi (ng)	Ntim (μL)
40	20

Cov ntaub so ntswg:

Exosome: 100 μL; 40 μg; >10^9 cov khoom me me/mL
Cov ntshav/ntshav: 5 mL
Cov tshuaj tua kab mob hauv lub hlwb: 100 mL
Cov zis: 100 mL
Cov qaub ncaug: 15 mL
Cov kua ntswg tawm: 15 mL
Cov kua dej hauv lub hlwb: 15 mL
Cov kua dej amniotic: 50 mL
Cov noob qes: 5 mL
Cov kua qaub: 5 ml
Ascites: 50 ml
Mis nyuj: 15 mL
Cov kua follicular: 15 mL
Cov kua txiv hmab txiv ntoo: 50 mL
Cov tshuaj tua kab mob hauv cov kab mob me me: 100 mL
Cov nqaij tsiaj: 3 g
Cov ntaub ntawm tsob nroj: 1 g

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

Tsim qauv kev sim

Kev xa khoom qauv

Kev rho tawm RNA

Kev tsim kho tsev qiv ntawv

Kev txheeb xyuas kab ke

Kev tshuaj xyuas cov ntaub ntawv

Cov kev pab cuam tom qab muag

Yav dhau los: BMKMANU S3000_Spatial Transcriptome

Tom ntej no: Exosomal mRNA/LncRNA/CircRNA Sequencing-Illumina

Kev tshawb fawb txog bioinformatics

● Kev tswj xyuas cov ntaub ntawv raw zoo
● kev faib tawm sRNA
● Kev sib phim rau ib qho genome siv ua piv txwv
● Kev txheeb xyuas cov miRNA uas paub thiab cov miRNA tshiab
● Kev tshuaj xyuas qhov sib txawv ntawm miRNA
● Kev piav qhia txog kev ua haujlwm ntawm cov hom phiaj miRNA

Kev txheeb xyuas miRNA: cov qauv thiab qhov tob