-
BMKMANU S3000_Spatial transcriptome
transcriptomics spatial na-eguzo n'ihu nke mmepụta sayensị, na-enye ndị nchọpụta ike ịbanye n'ime usoro okwu mkpụrụ ndụ mgbagwoju anya n'ime anụ ahụ ma na-echekwa ọnọdụ ha. N'etiti nyiwe dị iche iche, BMKGene ewepụtala BMKManu S3000 Spatial Transcriptome Chip, na-anya isi mkpebi siri ike nke 3.5µm, na-eru nso nso cell cell, na-enyere ntọala mkpebi ọkwa dị iche iche. Ihe mgbawa S3000, nke na-egosipụta ihe dị ka nde ntụpọ anọ, na-arụ ọrụ microwells nke nwere beads juru na nyocha njide kpuchiri oghere. A na-akwado ọba akwụkwọ cDNA, nke ejiri koodu barcode mejuo ya, site na mgbawa S3000 wee mee ya n'usoro n'elu ikpo okwu Illumina NovaSeq. Nchikota nke ihe nlele oghere na UMI na-eme ka o doo anya na izi ezi na nkọwapụta nke data ewepụtara. Ihe mgbawa BMKManu S3000 na-agbanwe agbanwe nke ukwuu, na-enye ntọala mkpebi ọkwa dị iche iche nke enwere ike ịhazi ya nke ọma na anụ ahụ dị iche iche yana ọkwa nkọwa achọrọ. Ngbanwe a na-edobe mgbawa ahụ dị ka nhọrọ pụtara ìhè maka ọmụmụ ihe gbasara transcriptomics dị iche iche, na-agba mbọ hụ na mkpokọta oghere nwere obere mkpọtụ. Ojiji teknụzụ nkewa cell na BMKManu S3000 na-enyere aka ikewapụta data ederede na ókèala nke mkpụrụ ndụ, na-ebute nyocha nke nwere nkọwa ndu kpọmkwem. Ọzọkwa, nkwubi okwu nke S3000 emelitere na-ebute ọnụ ọgụgụ dị elu nke mkpụrụ ndụ ihe nketa na UMI nke achọpụtara n'otu sel, na-eme ka nyocha nke ọma karịa nke usoro ntụgharị ohere na nchịkọta mkpụrụ ndụ.
-
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
-
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
