WO2024138523A1 - Procédé de séquençage transcriptomique spatio-temporel - Google Patents
Procédé de séquençage transcriptomique spatio-temporelInfo
- Publication number
- WO2024138523A1 WO2024138523A1 PCT/CN2022/143374 CN2022143374W WO2024138523A1 WO 2024138523 A1 WO2024138523 A1 WO 2024138523A1 CN 2022143374 W CN2022143374 W CN 2022143374W WO 2024138523 A1 WO2024138523 A1 WO 2024138523A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- sample
- cdna
- sequencing
- minutes
- spatiotemporal
- Prior art date
Links
- 238000012163 sequencing technique Methods 0.000 title claims abstract description 46
- 239000002299 complementary DNA Substances 0.000 claims abstract description 61
- 238000000034 method Methods 0.000 claims abstract description 45
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 16
- 239000000523 sample Substances 0.000 claims description 63
- 238000012408 PCR amplification Methods 0.000 claims description 18
- 238000010839 reverse transcription Methods 0.000 claims description 13
- 238000013518 transcription Methods 0.000 claims description 8
- 230000035897 transcription Effects 0.000 claims description 8
- 239000002773 nucleotide Substances 0.000 claims description 7
- 125000003729 nucleotide group Chemical group 0.000 claims description 7
- 238000009396 hybridization Methods 0.000 claims description 6
- 230000008823 permeabilization Effects 0.000 claims description 6
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 claims description 4
- 238000004132 cross linking Methods 0.000 claims description 4
- 230000002441 reversible effect Effects 0.000 claims description 4
- 238000012216 screening Methods 0.000 claims description 4
- 238000011529 RT qPCR Methods 0.000 claims description 2
- 238000007403 mPCR Methods 0.000 claims description 2
- 238000010008 shearing Methods 0.000 abstract description 12
- 238000005516 engineering process Methods 0.000 abstract description 11
- 238000011084 recovery Methods 0.000 abstract description 10
- 210000001519 tissue Anatomy 0.000 description 31
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 22
- 238000006243 chemical reaction Methods 0.000 description 21
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 18
- 239000000243 solution Substances 0.000 description 18
- 239000000047 product Substances 0.000 description 16
- 239000000203 mixture Substances 0.000 description 15
- 239000006228 supernatant Substances 0.000 description 14
- 239000000872 buffer Substances 0.000 description 13
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 12
- 239000003161 ribonuclease inhibitor Substances 0.000 description 12
- 108020004414 DNA Proteins 0.000 description 11
- 230000003321 amplification Effects 0.000 description 11
- 238000003199 nucleic acid amplification method Methods 0.000 description 11
- 239000007788 liquid Substances 0.000 description 9
- 239000011324 bead Substances 0.000 description 8
- 239000012188 paraffin wax Substances 0.000 description 8
- 210000004556 brain Anatomy 0.000 description 7
- 239000003153 chemical reaction reagent Substances 0.000 description 7
- 210000003734 kidney Anatomy 0.000 description 7
- 210000004072 lung Anatomy 0.000 description 7
- 230000002123 temporal effect Effects 0.000 description 6
- 102000053602 DNA Human genes 0.000 description 5
- 108090000790 Enzymes Proteins 0.000 description 5
- 102000004190 Enzymes Human genes 0.000 description 5
- 230000014509 gene expression Effects 0.000 description 5
- 238000007689 inspection Methods 0.000 description 5
- 238000000746 purification Methods 0.000 description 5
- 239000002096 quantum dot Substances 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 4
- 238000010586 diagram Methods 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 238000012545 processing Methods 0.000 description 4
- 108091028043 Nucleic acid sequence Proteins 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 238000009826 distribution Methods 0.000 description 3
- 239000012264 purified product Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 239000007984 Tris EDTA buffer Substances 0.000 description 2
- 239000002250 absorbent Substances 0.000 description 2
- 230000002745 absorbent Effects 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 230000000295 complement effect Effects 0.000 description 2
- 230000002596 correlated effect Effects 0.000 description 2
- 238000007405 data analysis Methods 0.000 description 2
- 238000005202 decontamination Methods 0.000 description 2
- 230000003588 decontaminative effect Effects 0.000 description 2
- 239000012634 fragment Substances 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 108020004999 messenger RNA Proteins 0.000 description 2
- 238000007363 ring formation reaction Methods 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 108010008286 DNA nucleotidylexotransferase Proteins 0.000 description 1
- 102100029764 DNA-directed DNA/RNA polymerase mu Human genes 0.000 description 1
- 101001121408 Homo sapiens L-amino-acid oxidase Proteins 0.000 description 1
- 102100026388 L-amino-acid oxidase Human genes 0.000 description 1
- 241000535824 Mastacembelocleidus bam Species 0.000 description 1
- 108091034117 Oligonucleotide Proteins 0.000 description 1
- 101100012902 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) FIG2 gene Proteins 0.000 description 1
- 238000003766 bioinformatics method Methods 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 230000008094 contradictory effect Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 238000012165 high-throughput sequencing Methods 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 238000003752 polymerase chain reaction Methods 0.000 description 1
- 238000003908 quality control method Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000007711 solidification Methods 0.000 description 1
- 230000008023 solidification Effects 0.000 description 1
- 210000001541 thymus gland Anatomy 0.000 description 1
- 238000011222 transcriptome analysis Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000009966 trimming Methods 0.000 description 1
Abstract
La présente invention concerne un procédé de séquençage transcriptomique spatiotemporel, comprenant les étapes suivantes : établissement de l'abondance de l'ADNc d'un échantillon à séquencer ; et réalisation d'une étape de séquençage transcriptomique spacio-temporelle sur ledit échantillon en fonction de l'abondance de l'ADNc, l'établissement de l'abondance de l'ADNc dudit échantillon comprenant une étape de séquençage transcriptomique pré-spatio-temporelle. Selon les modes de réalisation spécifiques de la présente divulgation, l'abondance de l'ADNc obtenue selon le procédé peut refléter avec précision la capacité de capture génique de la technique transcriptomique spatio-temporelle sur des coupes de tissus. Par comparaison avec les procédés existants, le procédé omet les étapes de récupération de l'ADNc et de cisaillement de l'ADNc, ce qui permet de réduire considérablement la durée de l'opération.
Publications (1)
Publication Number | Publication Date |
---|---|
WO2024138523A1 true WO2024138523A1 (fr) | 2024-07-04 |
Family
ID=
Similar Documents
Publication | Publication Date | Title |
---|---|---|
EP4087945B1 (fr) | Procédés de détermination d'un emplacement d'un acide nucléique cible dans un échantillon biologique | |
AU2019405940A1 (en) | Nuclease-based RNA depletion | |
JP7379418B2 (ja) | 腫瘍のディープシークエンシングプロファイリング | |
US8574832B2 (en) | Methods for preparing sequencing libraries | |
WO2019204560A1 (fr) | Procédé pour relier l'accessibilité de la chromatine et le transcriptome | |
US20070254305A1 (en) | Methods of whole genome or microarray expression profiling using nucleic acids prepared from formalin fixed paraffin embedded tissue | |
CN110129415B (zh) | 一种ngs建库分子接头及其制备方法和用途 | |
WO2016049878A1 (fr) | Procédé de test de filiation basé sur l'établissement de profils snp et application | |
CN112795654A (zh) | 用于生物体融合基因检测与融合丰度定量的方法及试剂盒 | |
CN108690873A (zh) | 21羟化酶基因突变的检测方法及检测试剂盒 | |
WO2016004548A1 (fr) | Procédé de pré-traitement de micro-quantités de tissus prélevés par biopsie clinique et enrobés de paraffne | |
AU2018367394A1 (en) | Method for making a cDNA library | |
WO2024138523A1 (fr) | Procédé de séquençage transcriptomique spatio-temporel | |
CN111705132A (zh) | 一种ddPCR检测肝癌预后标志物TP53 R249S的引物探针组、试剂盒和方法 | |
CN111020710A (zh) | 造血及淋巴组织肿瘤的ctDNA高通量检测 | |
CN114277092B (zh) | 基于纳米孔测序平台的rna病毒宏转录组建库方法及应用 | |
CN110273028A (zh) | 病毒整合型dna的富集方法、测序数据分析方法和装置 | |
CN115247203A (zh) | 用于构建空间转录组测序文库的试剂盒及cDNA文库的构建方法 | |
CN111534858B (zh) | 用于高通量测序的文库构建方法及高通量测序方法 | |
CN114250269A (zh) | 一种探针组合物、基于该探针组合物的二代测序文库及其应用 | |
CN112680794A (zh) | 一种应用于ngs平台的超微量核酸样本建库方法 | |
WO2024138670A1 (fr) | Séquence splint et son utilisation dans la capture d'acides nucléiques | |
CN113930487B (zh) | 一种新型多样本多片段dna甲基化检测方法 | |
CN116622806A (zh) | 一种提高rna病毒检出率的方法 | |
CN114507728A (zh) | 一种捕获引物及其应用 |