10x Genomics Visium Spatial Transcriptome
Atụmatụ
● Mkpebi: 100 µM
● Ogologo ntụpọ: 55 µM
● Ọnụ ọgụgụ nke ntụpọ: 4992
● Mpaghara ndọta: 6.5 x 6.5 mm
● A na-eju ebe ọ bụla akara koodu na primers nwere ngalaba anọ:
- poly(dT) ọdụ maka mRNA priming na cDNA njikọ
- Identifier Molecular pụrụ iche (UMI) iji mezie nhụsianya mmụba
- Barcode oghere
- Njikọ usoro nke akụkụ agụ 1 usoro primer
● H&E ntụpọ nke ngalaba
Uru
●Ọrụ nkwụsịtụ: na-ejikọta ahụmịhe niile na usoro dabere na nka, gụnyere ngalaba cryo, ncha, njikarịcha anụ ahụ, oghere oghere, nkwadebe ọba akwụkwọ, usoro na bioinformatics.
● Ndị otu ọkachamara nwere nkà: nwere ahụmahụ n'ihe karịrị ụdị anụ ahụ 250 na ụdị 100+ gụnyere mmadụ, òké, mammamal, azụ na osisi.
●Mmelite ozugbo na ọrụ ahụ dum: na njikwa zuru oke nke ọganihu nnwale.
●Ọkọlọtọ bioinformatics zuru oke:ngwugwu gụnyere nyocha 29 na ọnụ ọgụgụ 100+ dị elu.
●Nyocha na nleba anya data ahaziri: dị maka arịrịọ nyocha dị iche iche.
●Nyocha nkwonkwo nhọrọ yana usoro mRNA otu-cell
Nkọwapụta
| Ihe atụ chọrọ | Ụlọ akwụkwọ | Usoro usoro | akwadoro data | Njikwa ogo |
| Ihe nlere cryo agbakwunyere OCT, ihe nlele FFPE ( Dayameta kacha mma: ihe dịka 6x6x6 mm3) 3 ngọngọ kwa sample | 10X Visium cDNA ọba akwụkwọ | Ilumina PE150 | 50K PE na-agụ kwa ntụpọ (60Gb) | RIN>7 |
Maka nkọwa ndị ọzọ na ntụzịaka nkwadebe nlele na usoro ọrụ ọrụ, biko nweere onwe gị ịgwa aỌkachamara BMKGENE
Usoro ọrụ ọrụ
N'oge nkwadebe nlele, a na-eme nnwale mmịpụta RNA mbụ iji hụ na enwere ike nweta RNA dị elu.N'ọkwa nkwalite anụ ahụ, akụkụ ndị ahụ nwere ntụpọ ma na-ahụ anya ma na-emeziwanye ọnọdụ mRNA maka mwepụta anụ ahụ.A na-etinye usoro ahụ kachasị mma n'oge a na-arụ ụlọ ọba akwụkwọ, na-esote usoro na nyocha data.
Usoro ọrụ ọrụ zuru oke na-agụnye mmelite ozugbo yana nkwenye ndị ahịa iji nọgide na-enwe nzaghachi nzaghachi nzaghachi, na-eme ka mmezu ọrụ dị mma.
Ọ gụnyere nyocha ndị a:
Njikwa ogo data:
o Mpụta data na nkesa akara ogo
o Nchọpụta mkpụrụ ndụ ihe nketa kwa ntụpọ
o Mkpuchi anụ ahụ
Nleba anya n'ime:
o Gene ọgaranya
o Nchịkọta ntụpọ, gụnyere nyocha akụkụ dị ntakịrị
o Nyocha okwu dị iche n'etiti ụyọkọ: njirimara nke mkpụrụ ndụ ihe nrịbama
o Nkọwa ọrụ na ime ka mkpụrụ ndụ ihe nketa akara
Nyocha n'etiti otu
o Nchikota nke ntụpọ site na ma sample (eg. ọrịa na njikwa) na re-ụyọkọ
o Nchọpụta mkpụrụ ndụ ihe nrịbama maka ụyọkọ ọ bụla
o Nkọwa ọrụ na ime ka mkpụrụ ndụ ihe nketa akara
o Nkwuwapụta dị iche iche nke otu ụyọkọ n'etiti otu
Nleba anya n'ime
Nchịkọta ntụpọ
Nchọpụta mkpụrụ ndụ ihe nrịbama na nkesa oghere
Nyocha n'etiti otu
Ngwakọta data sitere na otu abụọ yana ụyọkọ ọzọ
Mkpụrụ ndụ ihe nrịbama nke ụyọkọ ọhụrụ
Nyochaa ọganihu ndị ọrụ transcriptomics nke BMKGene na-akwado site na 10X Visium N'ime akwụkwọ ndị a egosipụtara:
Chen, D. et al.(2023) 'mthl1, onye nwere ike Drosophila homologue nke mammalian adhesion GPCRs, na-etinye aka na mmeghachi omume antitumor na mkpụrụ ndụ oncogenic n'ime ijiji', Usoro nke National Academy of Sciences of the United States of America, 120(30), p.e2303462120.doi: /10.1073/pnas.2303462120
Chen, Y. et al.(2023) 'STEEL na-eme ka mkpebi dị elu dị elu nke data transcriptomic spatiotemporal', Briefings in Bioinformatics, 24(2), p. 1-10.doi: 10.1093/BIB/BBAD068.
Liu, C. et al.(2022) 'A spatiotemporal atlas of organogenesis na mmepe nke ifuru orchid', Nucleic Acids Research, 50(17), p. 9724-9737.doi: 10.1093/NAR/GKAC773.
Wang, J. et al.(2023) 'Ijikota Spatial Transcriptomics na Single-nucleus RNA Sequencing na-ekpughere Usoro ọgwụgwọ enwere ike maka Uterine Leiomyoma', International Journal of Biological Sciences, 19(8), p. 2515-2530.doi: 10.7150/IJBS.83510.
