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Tib- nuclei RNA Sequencing
Txoj kev loj hlob ntawm ib leeg-cell capture thiab kev cai tsev qiv ntawv tsim cov tswv yim, ua ke nrog high-throughput sequencing, tau revolutionized gene qhia kev kawm nyob rau hauv lub cell. Qhov kev rhuav tshem no tso cai rau kev sib sib zog nqus thiab kev tshuaj xyuas ntau dua ntawm cov cell complex, kov yeej cov kev txwv cuam tshuam nrog qhov nruab nrab ntawm cov noob qhia txog txhua lub hlwb thiab khaws cia qhov tseeb heterogeneity nyob rau hauv cov pejxeem. Thaum ib leeg-cell RNA sequencing (scRNA-seq) muaj qhov zoo tsis txaus ntseeg, nws ntsib cov teeb meem hauv qee cov ntaub so ntswg uas tsim cov kev ncua ntawm ib leeg ua pov thawj nyuaj thiab xav tau cov qauv tshiab. Ntawm BMKGene, peb hais txog qhov teeb meem no los ntawm kev muab ib lub nucleus RNA sequencing (snRNA-seq) siv lub xeev-of-the-art 10X Genomics Chromium thev naus laus zis. Txoj hauv kev no nthuav dav cov spectrum ntawm cov qauv ua kom zoo rau kev txheeb xyuas cov ntaub ntawv ntawm qib ib leeg.
Kev sib cais ntawm nuclei yog ua tiav los ntawm lub tshiab 10X Genomics Chromium nti, uas muaj yim-channel microfluidics system nrog ob txoj kev hla. Nyob rau hauv cov txheej txheem no, gel hlaws dai nrog barcodes, primers, enzymes, thiab ib lub nucleus yog encapsulated nyob rau hauv nanoliter-loj roj tee, tsim Gel Bead-in-Emulsion (GEM). Tom qab GEM tsim, cell lysis thiab barcode tso tawm tshwm sim hauv txhua GEM. Tom qab ntawd, mRNA molecules tau thim rov qab mus rau hauv cDNAs, suav nrog 10X barcodes thiab Cim Molecular Identifiers (UMIs). Cov cDNAs no tom qab ntawd raug rau cov qauv kev tsim kho tsev qiv ntawv, ua kom yooj yim rau kev tshawb nrhiav cov noob caj noob ces ntawm qib ib leeg.
Platform: 10 × Genomics Chromium thiab Illumina NovaSeq Platform
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10x Genomics Visium Spatial Transcriptome
Spatial transcriptomics yog ib qho kev siv thev naus laus zis uas tso cai rau cov kws tshawb fawb los tshawb xyuas cov qauv ntawm cov noob hauv cov ntaub so ntswg thaum khaws cia lawv cov ntsiab lus spatial. Ib lub platform muaj zog hauv qhov no yog 10x Genomics Visium ua ke nrog Illumina sequencing. Lub hauv paus ntsiab lus ntawm 10X Visium yog nyob ntawm ib qho tshwj xeeb nti nrog rau qhov chaw ntes uas tau muab tso rau hauv cov ntaub so ntswg. Qhov chaw ntes no muaj cov chaw barcoded, txhua qhov sib xws rau qhov chaw tshwj xeeb hauv cov ntaub so ntswg. Lub ntes RNA molecules los ntawm cov ntaub so ntswg yog tom qab ntawd sau nrog cov cim molecular identifiers (UMIs) thaum lub sijhawm rov qab cov txheej txheem. Cov chaw barcoded thiab UMIs pab kom meej spatial mapping thiab kom muaj nuj nqis ntawm cov noob qhia ntawm ib leeg-cell daws teeb meem. Kev sib xyaw ua ke ntawm spatial barcoded qauv thiab UMIs ua kom qhov tseeb thiab tshwj xeeb ntawm cov ntaub ntawv tsim tawm. Los ntawm kev siv thev naus laus zis Spatial Transcriptomics, cov kws tshawb fawb tuaj yeem nkag siab tob txog lub koom haum spatial ntawm cov hlwb thiab cov kev sib cuam tshuam ntawm cov molecular uas tshwm sim hauv cov ntaub so ntswg, muab kev nkag siab zoo rau cov txheej txheem hauv cov txheej txheem lom neeg hauv ntau qhov chaw, suav nrog oncology, neuroscience, kev loj hlob biology, immunology. , thiab kev tshawb fawb botanical.
Platform: 10X Genomics Visium thiab Illumina NovaSeq
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Full-Length mRNA Sequencing-Nanopore
Thaum NGS-raws li mRNA sequencing yog ntau yam cuab yeej rau kev txheeb xyuas cov noob qhia, nws qhov kev cia siab ntawm kev nyeem luv luv txwv nws txoj kev ua tau zoo hauv kev txheeb xyuas cov ntaub ntawv transcriptomic. Ntawm qhov tod tes, nanopore sequencing ntiav cov thev naus laus zis nyeem ntev, ua kom muaj kev sib luag ntawm cov ntawv sau tag nrho ntev mRNA. Txoj hauv kev no ua rau muaj kev tshawb nrhiav dav dav ntawm lwm txoj kev sib txuas, cov noob fusions, poly-adenylation, thiab qhov ntau ntawm mRNA isoforms.
Nanopore sequencing, ib txoj hauv kev uas tso siab rau nanopore ib leeg-molecule hluav taws xob lub sijhawm tiag tiag, muab cov txiaj ntsig hauv lub sijhawm. Kev coj los ntawm lub cev muaj zog proteins, ob-stranded DNA khi rau nanopore proteins embedded nyob rau hauv ib tug biofilm, unwinding raws li nws dhau los ntawm nanopore channel nyob rau hauv ib tug voltage sib txawv. Cov cim hluav taws xob tshwj xeeb tsim los ntawm cov hauv paus sib txawv ntawm DNA strand raug kuaj pom thiab cais tawm hauv lub sijhawm tiag tiag, ua kom muaj tseeb thiab txuas ntxiv nucleotide sequencing. Txoj hauv kev tshiab no kov yeej cov kev txwv luv luv thiab muab lub platform dynamic rau kev soj ntsuam genomic intricate, suav nrog cov kev tshawb fawb transcriptomic nyuaj, nrog rau cov txiaj ntsig tam sim ntawd.
Platform: Nanopore PromethION 48
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Tag nrho-ntev mRNA sequencing - PacBio
Thaum NGS-raws li mRNA sequencing yog ntau yam cuab yeej rau kev txheeb xyuas cov noob qhia, nws qhov kev cia siab ntawm kev nyeem luv luv txwv nws txoj kev siv hauv kev tshuaj ntsuam transcriptomic nyuaj. Ntawm qhov tod tes, PacBio sequencing (Iso-Seq) ntiav cov thev naus laus zis nyeem ntev, ua kom muaj kev sib luag ntawm cov ntawv sau ntev mRNA. Txoj hauv kev no ua rau muaj kev tshawb nrhiav dav dav ntawm lwm txoj kev sib txuas, noob fusions, thiab poly-adenylation. Txawm li cas los xij, muaj lwm txoj kev xaiv rau kev qhia cov noob kom muaj nuj nqis vim muaj cov ntaub ntawv xav tau ntau. PacBio sequencing thev naus laus zis tso siab rau ib leeg-molecule, real-time (SMRT) sequencing, muab qhov zoo sib xws hauv kev ntes tag nrho cov ntawv sau mRNA. Txoj hauv kev tshiab no suav nrog kev siv xoom-mode waveguides (ZMWs) thiab cov qhov dej microfabricated uas ua rau kev soj ntsuam lub sijhawm tiag tiag ntawm DNA polymerase kev ua haujlwm thaum lub sijhawm ua ntu zus. Hauv cov ZMWs no, PacBio's DNA polymerase synthesizes a complementary strand of DNA, generating long reading that span the tag nrho ntawm mRNA transcripts. PacBio ua haujlwm hauv Circular Consensus sequencing (CCS) hom txhim kho qhov tseeb los ntawm kev rov ua kom zoo li qub molecule. Cov ntawv nyeem HiFi generated muaj qhov tseeb piv rau NGS, ntxiv rau kev nthuav dav thiab txhim khu kev txheeb xyuas ntawm cov yam ntxwv ntawm cov ntaub ntawv nyuaj.
Platform: PacBio Sequel II; PacBio Revio: koj puas xav tau ntau tus thwjtim?
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Eukaryotic mRNA Sequencing-NGS
mRNA sequencing, ntau yam thev naus laus zis, txhawb nqa qhov kev qhia dav dav ntawm txhua mRNA cov ntawv teev lus hauv cov hlwb hauv cov xwm txheej tshwj xeeb. Nrog nws cov kev siv dav dav, cov cuab yeej txiav-ntug no nthuav tawm cov kab lus sib txawv ntawm cov noob caj noob ces, cov qauv noob, thiab cov txheej txheem molecular cuam tshuam nrog ntau cov txheej txheem lom neeg. Kev siv dav dav hauv kev tshawb fawb, kev kuaj mob, thiab kev txhim kho tshuaj, mRNA sequencing muab kev nkag siab rau hauv cov intricacies ntawm cellular dynamics thiab genetic regulation, sparking xav paub txog nws lub peev xwm nyob rau hauv ntau yam teb.
Platform: Illumina NovaSeq X; DNBSEQ-T7
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Non-Reference based mRNA Sequencing-NGS
mRNA sequencing txhawb nqa cov kev qhia dav dav ntawm txhua mRNA cov ntawv teev lus hauv cov hlwb hauv cov xwm txheej tshwj xeeb. Qhov kev siv thev naus laus zis no ua haujlwm zoo li cov cuab yeej muaj zog, nthuav tawm cov kab lus sib txawv ntawm cov noob caj noob ces, cov qauv noob, thiab cov txheej txheem molecular cuam tshuam nrog ntau yam txheej txheem lom neeg. Kev siv dav dav hauv kev tshawb fawb, kev kuaj mob, thiab kev tsim tshuaj, mRNA sequencing muab kev nkag siab rau hauv qhov tsis sib xws ntawm cellular dynamics thiab kev tswj caj ces.
Platform: Illumina NovaSeq X; DNBSEQ-T7
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Ntev Tsis-coding Sequencing-Illumina
Ntev tsis-coding RNAs (lncRNAs) ntev dua 200 nucleotides uas muaj peev xwm coding tsawg thiab yog cov ntsiab lus tseem ceeb hauv cov tsis-coding RNA. Pom muaj nyob rau hauv cov nucleus thiab cytoplasm, cov RNAs ua lub luag haujlwm tseem ceeb hauv kev tswj hwm ntawm epigenetic, transcriptional, thiab post-transcriptional regulation, underscoring lawv qhov tseem ceeb hauv shaping cellular thiab molecular txheej txheem. LncRNA sequencing yog ib qho cuab yeej muaj zog hauv Cell sib txawv, Ontogenesis, thiab Tib neeg kab mob.
Platform: Illumina NovaSeq
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Me RNA Sequencing-Illumina
Me RNA (sRNA) molecules, suav nrog microRNAs (miRNAs), me me cuam tshuam RNAs (siRNAs), thiab piwi-interacting RNAs (piRNAs). Ntawm cov no, miRNAs, nyob ib ncig ntawm 18-25 nucleotides ntev, tshwj xeeb tshaj yog tsim nyog rau lawv txoj haujlwm tseem ceeb ntawm kev tswj hwm hauv ntau cov txheej txheem ntawm tes. Nrog rau cov ntaub so ntswg tshwj xeeb thiab theem tshwj xeeb qhia cov qauv, miRNAs nthuav tawm kev txuag siab thoob plaws ntau hom.
Platform: Illumina NovaSeq
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CircRNA Sequencing-Illumina
Circular RNA sequencing (circRNA-seq) yog txhawm rau txheeb xyuas thiab txheeb xyuas RNAs, chav kawm ntawm RNA molecules uas tsim cov voj voog kaw vim tsis yog-canonical splicing txheej xwm, muab qhov RNA no nrog kev ruaj ntseg. Txawm hais tias qee qhov circRNAs tau pom tias ua raws li microRNA sponges, sequestering microRNAs thiab tiv thaiv lawv los ntawm kev tswj lawv lub hom phiaj mRNAs, lwm cov circRNAs yuav cuam tshuam nrog cov proteins, hloov cov noob qhia, lossis muaj lub luag haujlwm hauv cov txheej txheem ntawm tes. circRNA qhia kev tsom xam muab kev nkag siab txog kev tswj hwm lub luag haujlwm ntawm cov molecules thiab lawv qhov tseem ceeb hauv ntau yam txheej txheem ntawm tes, theem kev loj hlob, thiab kab mob, ua rau kom nkag siab tob txog qhov nyuaj ntawm RNA kev tswj hwm hauv cov ntsiab lus ntawm cov noob qhia.
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Tag nrho Transcriptome Sequencing - Illumina
Tag nrho transcriptome sequencing muab ib txoj hauv kev los qhia txog kev sib txawv RNA molecules, encompassing coding (mRNA) thiab non-coding RNAs (lncRNA, circRNA, thiab miRNA). Cov txheej txheem no ntes tag nrho cov ntawv teev lus ntawm cov hlwb tshwj xeeb ntawm ib lub sijhawm, tso cai rau kev nkag siab zoo ntawm cov txheej txheem cellular. Kuj tseem hu ua "tag nrho RNA sequencing," nws lub hom phiaj los nthuav tawm cov kev tswj hwm kev sib txuas lus sib txuas ntawm qib kev sau ntawv, ua kom muaj kev soj ntsuam tob xws li kev sib tw endogenous RNA (ceRNA) thiab sib koom RNA tsom xam. Qhov no yog thawj kauj ruam ntawm kev ua haujlwm zoo, tshwj xeeb tshaj yog nyob rau hauv kev nthuav tawm cov kev tswj hwm kev sib koom tes uas cuam tshuam nrog circRNA-miRNA-mRNA-based ceRNA kev sib cuam tshuam.
