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Otu-nuclei RNA Sequencing
Mmepe nke ijide otu cell na usoro iwu ụlọ ọba akwụkwọ ahaziri ahazi, yana usoro ntinye dị elu, agbanweela ọmụmụ ihe ọmụmụ mkpụrụ ndụ ihe nketa na ọkwa cell. Ọganihu a na-enye ohere maka nyocha miri emi na nke sara mbara nke ọnụ ọgụgụ sel ndị dị mgbagwoju anya, na-emeri njedebe jikọtara ya na nkezi mkpụrụ ndụ ihe nketa n'elu sel niile na ichekwa ezi ihe dị iche iche n'ime ndị a. Ọ bụ ezie na usoro RNA otu cell (scRNA-seq) nwere uru a na-apụghị ịgbagha agbagha, ọ na-ezute ihe ịma aka na ụfọdụ anụ ahụ ebe imepụta nkwusioru otu cell na-esi ike ma chọọ nlele ọhụrụ. Na BMKGene, anyị na-edozi nsogbu a site na ịnye usoro RNA otu-nucleus (snRNA-seq) site na iji teknụzụ 10X Genomics Chromium ọgbara ọhụrụ. Usoro a na-agbasawanye ụdị nlele nke enwere ike iji nyocha transcriptome na ọkwa otu cell.
A na-enweta ikewapụ nuclei site na mgbawa 10X Genomics Chromium ọhụrụ, na-egosipụta sistemụ microfluidics ọwa asatọ nwere ngafe abụọ. N'ime usoro a, gel beads gụnyere barcodes, primers, enzymes, na otu nucleus na-agbanye n'ime mmanụ na-anaoliter, na-eme Gel Bead-in-Emulsion (GEM). Na-esote nhazi GEM, cell lysis na ntọhapụ koodu na-eme n'ime GEM ọ bụla. Na-esote, ụmụ irighiri mRNA na-enweta ntụgharị ntụgharị n'ime cDNAs, na-etinye koodu barcode 10X na Ihe nchọpụta Molecular Pụrụ Iche (UMIs). A na-edokwa cDNA ndị a n'iwu ụlọ ọba akwụkwọ usoro nhazi ọkọlọtọ, na-eme ka nyocha siri ike na nke zuru oke nke profaịlụ mkpụrụ ndụ ihe nketa na ọkwa otu cell.
Platform: 10× Genomics Chromium na Illumina NovaSeq Platform
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10x Genomics Visium Spatial Transcriptome
transcriptomics spatial bụ nkà na ụzụ na-emepe emepe nke na-enye ndị nchọpụta ohere inyocha usoro okwu mkpụrụ ndụ ihe nketa n'ime anụ ahụ ma na-echekwa ọnọdụ ha. Otu ikpo okwu siri ike na ngalaba a bụ 10x Genomics Visium yana usoro Illumina. Ụkpụrụ nke 10X Visium dabere na mgbawa pụrụ iche nwere mpaghara ejidere ahaziri ebe etinye akụkụ anụ ahụ. Mpaghara njide a nwere ntụpọ akara, nke ọ bụla dabara na ọnọdụ oghere pụrụ iche n'ime anụ ahụ. A na-edezi ụmụ irighiri ihe RNA ndị ahụ e weghaara si n'anụ ahụ nwere ihe nchọpụta molekụla pụrụ iche (UMIs) n'oge usoro ntụgharị ntụgharị. Ebe akara koodu na UMI ndị a na-eme ka eserese oghere na ọnụọgụ mkpụrụ ndụ ihe nketa na mkpebi otu cell. Nchikota nke ihe nlele oghere na UMI na-eme ka o doo anya na izi ezi na nkọwapụta nke data ewepụtara. Site n'iji teknụzụ Spatial Transcriptomics a, ndị nchọpụta nwere ike nweta nghọta miri emi banyere nhazi oghere nke mkpụrụ ndụ yana mmekọrịta dị mgbagwoju anya na-eme n'ime anụ ahụ, na-enye nghọta bara uru n'ime usoro ndị na-ebute usoro ndu n'ọtụtụ mpaghara, gụnyere oncology, neuroscience, bayoloji mmepe, immunology. , na ọmụmụ ihe gbasara ahịhịa.
Platform: 10X Genomics Visium na Illumina NovaSeq
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Usoro mRNA dị ogologo-Nanopore
Ọ bụ ezie na usoro mRNA dabeere na NGS bụ ngwá ọrụ dị iche iche maka ịkọwapụta mkpụrụ ndụ ihe nketa, ịdabere na mkpirisi agụ na-egbochi ịdị irè ya na nyocha transcript dị mgbagwoju anya. N'aka nke ọzọ, usoro nanopore na-eji teknụzụ na-agụ ogologo oge, na-eme ka usoro nke transcript mRNA zuru oke. Usoro a na-eme ka nyocha zuru oke nke splicing ọzọ, njikọ mkpụrụ ndụ ihe nketa, poly-adenylation, na ọnụọgụ nke isoforms mRNA.
Usoro Nanopore, usoro na-adabere na nanopore otu molecule ozugbo akara ọkụ eletrik, na-enye nsonaazụ na oge. N'ịbụ ndị na-eduzi proteins moto, DNA nwere eriri abụọ na-ejikọta na protein nanopore agbakwunyere na biofilm, na-atụgharị ka ọ na-agafe na ọwa nanopore n'okpuru ọdịiche voltaji. A na-achọpụta akara ọkụ eletrik pụrụ iche nke ntọala dị iche iche na-emepụta na eriri DNA na nkewa ozugbo, na-eme ka usoro nucleotide ziri ezi na na-aga n'ihu. Ụzọ ọhụrụ a na-emeri njedebe nke ọgụgụ dị mkpirikpi ma na-enye ikpo okwu siri ike maka nyocha nke genomic dị mgbagwoju anya, gụnyere nchọpụta transcript dị mgbagwoju anya, yana nsonaazụ ozugbo.
Platform: Nanopore PromethION 48
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Usoro mRNA zuru oke -PacBio
Ọ bụ ezie na usoro mRNA dabere na NGS bụ ngwá ọrụ dị iche iche maka ịkọwapụta mkpụrụ ndụ ihe nketa, ịdabere na mkpirisi agụ na-egbochi ojiji ya na nyocha transcript dị mgbagwoju anya. N'aka nke ọzọ, PacBio sequencing (Iso-Seq) na-eji teknụzụ na-agụ ogologo oge, na-eme ka usoro nke transcript mRNA zuru oke. Usoro a na-eme ka nyocha zuru oke nke splicing ọzọ, njikọ mkpụrụ ndụ ihe nketa, na poly-adenylation. Agbanyeghị, enwere nhọrọ ndị ọzọ maka ọnụọgụ mkpụrụ ndụ ihe nketa n'ihi oke data achọrọ. Teknụzụ usoro PacBio na-adabere na usoro otu-molecule, ezigbo oge (SMRT), na-enye uru dị iche iche n'inweta ndegharị mRNA zuru oke. Uzo ohuru ohuru a gunyere iji uzo ebili mmiri efu (ZMW) na olulu mmiri microfabricated nke na-enyere aka nleba anya nke DNA polymerase n'oge usoro. N'ime ZMW ndị a, PacBio's DNA polymerase na-emepụta eriri DNA na-emekọ ihe, na-ewepụta ogologo ọgụgụ nke na-agafe mkpokọta mRNA. Ịrụ ọrụ PacBio na usoro nkwenye nkwekọrịta okirikiri (CCS) na-akwalite izi ezi site n'ịhazi otu molekul ugboro ugboro. Ọgụgụ HiFi emepụtara nwere izi ezi nke atụnyere NGS, na-enye aka n'ihu na nyocha zuru oke na ntụkwasị obi nke njirimara transcriptomic siri ike.
Platform: PacBio Sequel II; PacBio Revio
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Usoro nke Eukaryotic mRNA-NGS
usoro mRNA, teknụzụ na-agbanwe agbanwe, na-enye ike ịkọwapụta mkpokọta mRNA niile n'ime sel n'okpuru ọnọdụ akọwapụtara. Site na ngwa ya dị ukwuu, ngwá ọrụ a na-egbu egbu na-ekpughe profaịlụ mkpụrụ ndụ dị mgbagwoju anya, usoro mkpụrụ ndụ ihe nketa, na usoro molekụla jikọtara ya na usoro ndu dị iche iche. N'ịbụ nke anakwere na nyocha bụ isi, nchọpụta ụlọ ọgwụ, na mmepe ọgwụ, usoro mRNA na-enye nghọta n'ime mgbagwoju anya nke usoro mkpụrụ ndụ ihe nketa na ụkpụrụ mkpụrụ ndụ ihe nketa, na-akpalite ọchịchọ ịmata ikike ya n'akụkụ dị iche iche.
Platform: Illumina NovaSeq X; DNBSEQ-T7
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Usoro mRNA na-abụghị akwụkwọ ntụaka-NGS
Usoro mRNA na-enye ike ịkọwapụta mkpokọta mRNA niile n'ime sel n'okpuru ọnọdụ akọwapụtara. Teknụzụ ọgbara ọhụrụ a na-arụ ọrụ dị ka ngwa ọrụ siri ike, na-ekpughe profaịlụ mkpụrụ ndụ dị mgbagwoju anya, usoro mkpụrụ ndụ ihe nketa, na usoro mkpụrụ ndụ jikọtara ya na usoro ndu dị iche iche. N'ịbụ nke anakwere na nyocha dị mkpa, nchọpụta ụlọ ọgwụ, na mmepe ọgwụ, usoro mRNA na-enye nghọta n'ime mgbagwoju anya nke ike ndụ ekwentị na ụkpụrụ mkpụrụ ndụ ihe nketa.
Platform: Illumina NovaSeq X; DNBSEQ-T7
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Ogologo usoro enweghị koodu-Ilumina
Ogologo RNA ndị na-abụghị koodu (lncRNAs) dị ogologo karịa 200 nucleotides nwere ike itinye koodu ma bụrụ ihe dị mkpa n'ime RNA na-abụghị koodu. Achọpụtara na nucleus na cytoplasm, ndị RNA ndị a na-arụ ọrụ dị mkpa na epigenetic, transcriptional, na post-transcriptional regulation, na-egosi mkpa ha dị na ịkpụzi usoro cellular na molecular. Usoro LncRNA bụ ngwa ọrụ siri ike na ọdịiche nke mkpụrụ ndụ, Ontogenesis na ọrịa mmadụ.
Platform: Illumina NovaSeq
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Obere RNA Sequencing-Ilumina
Obere ụmụ irighiri RNA (sRNA), gụnyere microRNAs (miRNAs), obere RNA (siRNAs), na RNA piwi na-emekọrịta ihe (piRNA). N'ime ndị a, miRNA, gburugburu 18-25 nucleotides ogologo, bụ ihe kwesịrị ịrịba ama maka ọrụ nchịkwa ha dị mkpa na usoro cellular dị iche iche. Site na usoro ngosipụta nke anụ ahụ na nke akọwapụtara nke ọma, miRNA na-egosipụta nchekwa dị elu n'ofe ụdị dị iche iche.
Platform: Illumina NovaSeq
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Usoro nke CircRNA-Ilumina
okirikiri RNA sequencing (circRNA-seq) bụ profaịlụ na nyochaa okirikiri RNA, otu klas nke RNA molekul na-etolite mechiri emechi n'ihi na-abụghị nke canonical splicing omume, na-enye RNA a nkwụsi ike. Ọ bụ ezie na egosila ụfọdụ circRNA ka ha na-eme ihe dị ka sponge microRNA, na-achọgharị microRNA ma na-egbochi ha ịhazi mRNA ndị ha lekwasịrị anya, circRNA ndị ọzọ nwere ike imekọrịta ihe na protein, gbanwee mkpụrụ ndụ ihe nketa, ma ọ bụ nwee ọrụ na usoro cellular. Nyocha okwu circRNA na-enye nghọta n'ime ọrụ nhazi nke ụmụ irighiri ihe ndị a na mkpa ha dị na usoro cellular dị iche iche, ọkwa mmepe, na ọnọdụ ọrịa, na-enye aka n'ịghọta miri emi nke mgbagwoju anya nke ụkpụrụ RNA na ọnọdụ nke okwu mkpụrụ ndụ ihe nketa.
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Usoro transcriptome niile – Illumina
Usoro transcriptome niile na-enye ụzọ zuru oke maka ịkọwapụta mkpụrụ ndụ RNA dị iche iche, gụnyere koodu (mRNA) na RNA ndị na-abụghị koodu (lncRNA, circRNA, na miRNA). Usoro a na-ewepụta transcriptome nke mkpụrụ ndụ akọwapụtara n'otu oge, na-enye ohere maka nghọta zuru oke nke usoro cellular. Amarakwa dị ka “ngụkọta usoro RNA,” ọ chọrọ ikpughe netwọọdụ usoro iwu dị mgbagwoju anya na ọkwa transcriptome, na-enye nyocha miri emi dị ka asọmpi endogenous RNA (ceRNA) na nyocha RNA nkwonkwo. Nke a na-egosi nzọụkwụ mbụ n'ịkọwapụta ọrụ, ọkachasị n'iwepụta netwọkụ nchịkwa metụtara mmekọrịta ceRNA dabere na circRNA-miRNA-mRNA.
