¡Ø º¯°æ»çÇ×
‘SMART-Seq
® mRNA LP’¿Í ‘SMART-Seq
® mRNA’´Â °¢°¢ ‘SMART-Seq
® v4 PLUS’¿Í ‘SMART-Seq
® v4 Ultra Low Input RNA Kit for Sequencing’ÀÇ µ¿ÀÏÁ¦Ç°À¸·Î, Á¦Ç°¸í º¯°æ ¹× Á¦Ç°°£ ±¸¼ºÇ° Á¾·ù¸¦ ÅëÀÏÇÏ°í ¹ÝÀÀ ´ç ´Ü°¡¸¦ ÀÎÇÏÇÏ¿´½À´Ï´Ù
ÀÚ¼¼ÇÑ ³»¿ëÀº ´ÙÀ½ ¾È³»ÆäÀÌÁö¸¦ Âü°íÇϼ¼¿ä! Á¦Ç° ¼±ÅÃÀÌ ½¬¿öÁý´Ï´Ù.
>> RNA-Seq
Á¦Ç°¸íÀÌ ÀÌ·¸°Ô ¹Ù²ò´Ï´Ù
¡Ø ¾÷µ¥ÀÌÆ® ³»¿ë
¾Æ·¡ÀÇ ¾÷µ¥ÀÌÆ® »çÇ×Àº ½ÇÇè°úÁ¤À̳ª Á¦Ç° È¿À²¿¡ ¿µÇâÀÌ ¾ø½À´Ï´Ù.
- Total RNA Input range°¡ 10 pg-10 ng¿¡¼ 10 pg-100 ng·Î È®Àå
- SeqAmp PCR buffer´ë½Å SeqAmp CB PCR buffer·Î Á¦°øµÇ¾î, DNA cleanup °úÁ¤ÀÇ bead pelleting°ú resuspension Çâ»ó
- Á¦Ç°»ç¿ë À¯¿¬¼ºÀ» À§ÇØ index°¡ kit¿¡ Æ÷ÇԵǾî ÀÖÁö ¾ÊÀ¸¸ç, º°µµ·Î ±¸¸Å°¡ °¡´É >> ȣȯµÇ´Â Unique Dual Index kits
- Afa I, 10X Afa I Buffer, 0.1% BSA ¹ÌÁ¦°ø (½ÇÇè°úÁ¤ Áß ÇÊ¿äÇÏÁö ¾ÊÀ½)
- 10 pg ¼öÁØÀÇ ±Ø¼Ò·® total RNA¿¡µµ Àû¿ë °¡´É
- Unparalleled sensitivity and reproducibility
- Single-tube workflows - cDNA ÇÕ¼ººÎÅÍ library Á¦ÀÛ °úÁ¤À» °¢°¢ single tube¿¡¼ ÁøÇàÇÒ ¼ö ÀÖ¾î, »ùÇà ¼Õ½ÇÀ̳ª ¿À¿° µî ½ÇÇè °úÁ¤¿¡¼ ¹ß»ýÇÒ ¼ö ÀÖ´Â ¿À·ù ÃÖ¼ÒÈ
- High-quality RNA-seq data - total RNA·ÎºÎÅÍ °¡Àå ¸¹Àº ¼öÀÇ À¯ÀüÀÚ ºÐ¼®
- Increase your sequencing power– ¸¹Àº ¼öÀÇ »ùÇÃÀ» Çѹø¿¡ ºÐ¼® ½Ã, Unique dual index (UDI)¸¦ »ç¿ëÇÏ¿© index hoping ¾øÀÌ multiplexing ºÐ¼®ÀÌ °¡´É
- High library yield - ³ôÀº yieldÀÇ library¸¦ Á¦ÀÛÇÒ ¼ö ÀÖ¾î,NovaSeq™ system°ú °°Àº high-throughput sequencer¿¡ Àû¿ëÇϰųª, ´ÙȸºÐ¼®ÀÌ °¡´É
Á¦Ç°¼³¸í
SMART-Seq® mRNA Á¦Ç°À» ÀÌ¿ëÇϸé, 10 pg–100 ng total RNA ¶Ç´Â 1-1,000°³ÀÇ ¼¼Æ÷¿¡¼ RNA ÃßÃâ °úÁ¤ ¾øÀÌ, oligo(dT) primer¸¦ ÀÌ¿ëÇÑ °íǰÁúÀÇ full-length cDNA¸¦ Á¦Á¶ÇÒ ¼ö ÀÖ´Ù.
º» Á¦Ç°Àº SMART® (Switching Mechanism at 5' End of RNA Template) ±â¼ú¿¡ Locked Nucleic Acid (LNA) ±â¼úÀ» Á¢¸ñÇÔÀ¸·Î½á template switchingÀÇ È¿À²À» ´õ¿í Çâ»ó½ÃÄ×´Ù (±×¸²1). µû¶ó¼, ÀÌÀü ¼¼´ëÀÇ Á¦Ç°º¸´Ù ´õ¿í ¸¹Àº ¼öÀÇ À¯ÀüÀÚ¸¦ °ËÃâÇÒ ¼ö ÀÖ¾î Àü¹ÝÀûÀÎ Àü»çü ¿¬±¸ (full transcriptome analysis)¿¡ ÀÖ¾î ¸Å¿ì À¯¿ëÇÏ´Ù. SMART-Seq® mRNA´Â ³ôÀº ÀçÇö¼º°ú gene-body coverage »Ó ¾Æ´Ï¶ó GC-rich Àü»çüµµ Á¤È®ÇÏ°Ô cDNA·Î ÇÕ¼ºÇÒ ¼ö ÀÖ´Ù.
SMART-Seq mRNA® LP¿¡ Æ÷ÇԵǾî ÀÖ´Â library preparation kit´Â enzymatic fragmentation¿Í stem-loop adaptor·Î 2½Ã°£ ³»¿¡ single tube¿¡¼ º°µµÀÇ purification °úÁ¤ ¾øÀÌ NGS library±îÁö Á¦ÀÛÇÒ ¼ö ÀÖ´Ù. µû¶ó¼ »ùÇà ¼Õ½ÇÀ̳ª ¿À¿° µî ½ÇÇè °úÁ¤¿¡¼ ¹ß»ýÇÒ ¼ö ÀÖ´Â ¿À·ù¸¦ ÃÖ¼ÒÈÇÒ ¼ö ÀÖ´Ù.
ÀÌ °úÁ¤À» ÅëÇØ Á¦ÀÛµÈ library´Â SMART-Seq2 µî ±âÁ¸ ¹æ¹ý ¹× ´ç»ç ±âÁ¸ Á¦Ç° (SMART-Seq® v3)¿¡ ºñÇØ ÀçÇö¼ºÀÌ ³ô°í, ÈξÀ ¿ì¼öÇÒ »Ó ¾Æ´Ï¶ó, ³ôÀº yield·Î Á¦À۵Ǿî high-throughput sequencer¿¡ Àû¿ëÇϰųª, ÇѹøÀÇ ¹ÝÀÀÀ¸·Î ¿©·¯ Â÷·Ê ¹Ýº¹ ºÐ¼®ÀÌ °¡´ÉÇÏ´Ù. ¶ÇÇÑ ±âÁ¸ Á¦Ç°ÀÎ SMART-Seq® v4 Ultra® Low Input RNA Kit for Sequencing, SMART-Seq® v4 PLUS Kit ¸¸ÅÀÇ ³ôÀº °¨µµ¸¦ °¡Áø´Ù.
SMART-Seq mRNA LP (with UMIs)´Â SMART® ¹æ¹ýÀ¸·Î oligo¿¡ Unique Molecular Identifiers (UMI)s¸¦ ºÎ°¡ÇÔÀ¸·Î½á PCR errors, duplicates, amplification biases¸¦ º¸Á¤ÇÑ´Ù.
SMART-Seq mRNA® LP, SMART-Seq® mRNA LP (with UMIs)´Â »ùÇÿ¡¼ cDNA ÇÕ¼º, library prep¹× ¹«·á Cogent NGS »ý¹°Á¤º¸ÇÐ ÆÄÀÌÇÁ¶óÀÎÀ» ÅëÇÑ µ¥ÀÌÅÍ ºÐ¼®¿¡ À̸£±â±îÁö end-to-end solutionÀ» Á¦°øÇÑ´Ù.
Cogent NGS ¼ÒÇÁÆ®¿þ¾î´Â ¾Æ·¡ ÆäÀÌÁö¿¡¼ ´Ù¿î·Îµå ¹ÞÀ» ¼ö ÀÖ´Ù.
Cogent NGS Discovery Software(takarabio.com)
Cogent NGS Analysis Pipeline(takarabio.com)

±×¸² 1. SMART±â¼úÀ» ÀÌ¿ëÇÑ cDNA ÇÕ¼º °úÁ¤

±×¸²2. SMART-Seq mRNA® LP¸¦ ÀÌ¿ëÇÑ Single tube library prep ¿öÅ©Ç÷οì
Library prep °úÁ¤¿¡ UDI (Unique Dual Index) kit (Code 634752 ¿Ü, º°µµ ÆÇ¸Å)¸¦ ÇÔ²² »ç¿ëÇϸé ÃÖ´ë 384°³ÀÇ multiplexed Illumina-ready sequencing libraries¸¦ »ý¼ºÇÒ ¼ö ÀÖ´Ù. º» Á¦Ç°Àº high-throughput »ùÇà 󸮿ë liquid handlers¸¦ À§ÇÑ »ç¿ë·® ÃÖÀûÈ (miniaturization) ¹× ÀÚµ¿È (automation) ¿¡µµ ȣȯµÈ´Ù. Degraded samples¿¡ Àû¿ëÇÒ ¼ö ÀÖ´Â random priming ¹æ¹ýÀ» ¼±È£ÇÒ °æ¿ì, Stranded total RNA-Seq kit (Code 634485 ¿Ü)¸¦ ±ÇÀåÇÑ´Ù.

±×¸²3. Unique Molecular Identifier (UMI)¸¦ žÀçÇÑ SMART® ¹ýÀÇ cDNA ÇÕ¼º ¿öÅ©Ç÷οì SMART-Seq® mRNA LP (with UMIs) Á¦Ç°À» »ç¿ë½Ã, UMIs ºÎ°¡·Î Á¤È®µµ¿Í ½Å·Úµµ¸¦ ´õ ³ôÀÏ ¼ö ÀÖ´Ù.

±×¸²4. Single-tube ¹ÝÀÀÀ» ÅëÇÑ °£ÆíÇÑ library perp

±×¸²5. CongentDS À¥ ÀÎÅÍÆäÀ̽ºÀÇ UMAP ºÐ¼® Â÷Æ® ¿¹½Ã. Chart display¸¦ ¼öÁ¤Çϱâ À§ÇÑ user controlÀÌ scatterplot °á°ú¿Í ÇÔ²² º¸¿©Áø´Ù.
Àû¿ë
±Ø¼Ò·® »ùÇ÷κÎÅÍ full-length transcriptome ºÐ¼® (transcript isoforms, gene fusions, point mutations µî)À» À§ÇØ, cDNA ÇÕ¼º °úÁ¤¿¡¼ »ç¿ëÇÒ ¼ö ÀÖ´Ù (SMART-Seq mRNA ÀÌ¿ë).
SMART-Seq® mRNA·Î ÇÕ¼ºÇÑ cDNA´Â SMART-Seq® Library Prep kit (SMART-Seq® mRNA LP ³» Æ÷ÇÔ, º°µµ ±¸¸Å ºÒ°¡)¸¦ ÀÌ¿ëÇÏ¿© Illumina® ºÐ¼®¿ë sequencing library·Î Á¦ÀÛÇÒ ¼ö ÀÖ´Ù.
Components & Storage Conditions
°¢ Á¦Ç° ±¸¼º¹°°ú º¸°üÁ¶°ÇÀº Certificate of Analysis¸¦ ÂüÁ¶ÇϽʽÿÀ.
Citation
‘SMART-Seq® v4 PLUS’¿Í ‘SMART-Seq® v4 Ultra Low Input RNA Kit for Sequencing’ Á¦Ç°Àº °¢°¢ ‘SMART-Seq® mRNA LP’¿Í ‘SMART-Seq® mRNA’À¸·Î Á¦Ç°¸íÀÌ º¯°æµÇ¾ú½À´Ï´Ù. CitationÀÌ µ¿ÀÏÇÕ´Ï´Ù.
Multi-omic analyses of changes in the tumor microenvironment of pancreatic adenocarcinoma following neoadjuvant treatment with anti-PD-1 therapy
Cancer cell | 2022 Oct 27 | PubMed ID: 36306792 | Read Article
PD-1 -cis IL-2R agonism yields better effectors from stem-like CD8 + T cells
Nature | 2022 Sep 28 | PubMed ID: 36171284 | Read Article
An amygdala circuit that suppresses social engagement
Nature | 2022 Jul 04 | PubMed ID: 33790466 | Read Article
An engineered IL-2 partial agonist promotes CD8 + T cell stemness
Nature | 2022 Jun 07 | PubMed ID: 34526724 | Read Article
The spatial landscape of progression and immunoediting in primary melanoma at single cell resolution
Cancer discovery | 2022 Jun 02 | PubMed ID: 35404441 | Read Article
MCM complexes are barriers that restrict cohesin-mediated loop extrusion
Nature | 2022 May 18 | PubMed ID: 35585235 | Read Article
Extricating human tumour immune alterations from tissue inflammation
Nature | 2022 May 11 | PubMed ID: 35545675 | Read Article
Anatomic position determines oncogenic specificity in melanoma
Nature | 2022 Apr 01 | PubMed ID: 35355015 | Read Article
Corticosterone inhibits Gas6 to govern hair follicle stem cell quiescence
Nature | 2022 Mar 15 | PubMed ID: 33790465 | Read Article
An autoimmune stem-like CD8 T cell population drives type 1 diabetes
Nature | 2022 Feb 01 | PubMed ID: 34847567 | Read Article
Tissue-resident macrophages provide a pro-tumorigenic niche to early NSCLC cells.
Nature | 2022 Jan 11 | PubMed ID: 34135508 | Read Article
Exercise plasma boosts memory and dampens brain inflammation via clusterin
Nature | 2021 Dec 08 | PubMed ID: 34880498 | Read Article
Fate mapping of single NK cells identifies a type 1 innate lymphoid-like lineage that bridges innate and adaptive recognition of viral infection
Immunity | 2021 Oct 12 | PubMed ID: 34437840 | Read Article
Human neocortical expansion involves glutamatergic neuron diversification
Nature | 2021 Oct 06 | PubMed ID: 34616067 | Read Article
A transcriptomic and epigenomic cell atlas of the mouse primary motor cortex
Nature | 2021 Oct 06 | PubMed ID: 34616066 | Read Article
Comparative cellular analysis of motor cortex in human, marmoset and mouse
Nature | 2021 Oct 06 | PubMed ID: 34616062 | Read Article
Regulation of Prefrontal Patterning and Connectivity by Retinoic Acid
Nature | 2021 Oct 01 | PubMed ID: 34599305 | Read Article
Distinct developmental pathways from blood monocytes generate human lung macrophage diversity.
Immunity | 2021 Sep 07 | PubMed ID: 33382972 | Read Article
Adult neural stem cell activation in mice is regulated by the day/night cycle and intracellular calcium dynamics.
Cell | 2021 Aug 24 | PubMed ID: 33482084 | Read Article
Genetic insights into biological mechanisms governing human ovarian ageing
Nature | 2021 Aug 01 | PubMed ID: 34349265 | Read Article
The Identity of Human Tissue-Emigrant CD8 + T Cells.
Cell | 2021 May 18 | PubMed ID: 33306960 | Read Article
Notch4 signaling limits regulatory T-cell-mediated tissue repair and promotes severe lung inflammation in viral infections
Immunity | 2021 Apr 28 | PubMed ID: 33915108 | Read Article
Impaired Kupffer Cell Self-Renewal Alters the Liver Response to Lipid Overload during Non-alcoholic Steatohepatitis.
Immunity | 2021 Apr 22 | PubMed ID: 32562600 | Read Article
H2AK119ub1 guides maternal inheritance and zygotic deposition of H3K27me3 in mouse embryos.
Nature genetics | 2021 Apr 20 | PubMed ID: 33821003 | Read Article
CAR directs T cell adaptation to bile acids in the small intestine
Nature | 2021 Apr 07 | PubMed ID: 33828301 | Read Article
Osteoclasts recycle via osteomorphs during RANKL-stimulated bone resorption
Cell | 2021 Mar 04 | PubMed ID: 33636130 | Read Article
Dynamic Regulation of Tfh Selection During the Germinal Center Reaction
Nature | 2021 Feb 03 | PubMed ID: 33536617 | Read Article
Arterialization requires the timely suppression of cell growth
Nature | 2021 Jan 26 | PubMed ID: 33299176 | Read Article
Microbiota-Induced Type I Interferons Instruct a Poised Basal State of Dendritic Cells.
Cell | 2020 Dec 16 | PubMed ID: 32380006 | Read Article
SWI/SNF Complex Mutations Promote Thyroid Tumor Progression and Insensitivity to Redifferentiation Therapies
Cancer Discovery | 2020 Dec 14 | Read Article
>> ³í¹® ´õ º¸±â (¿¬°áµÇ´Â ÆäÀÌÁö ÇÏ´ÜÀ» Âü°íÇØÁÖ¼¼¿ä)