- Robust, full-length chemistry - ±âÁ¸ÀÇ ¸ðµç single cell full-length ºÐ¼®¹ýÀ» ¶Ù¾î³Ñ´Â ¼º´É
- Easy, plate-based workflow- FACS µîÀ¸·Î ºÐ¸®ÇÑ single cell¿¡ ¹Ù·Î Àû¿ë °¡´É
- 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
- Unparalleled sensitivity and reproducibility- ³ôÀº À¯ÀüÀÚ °ËÃâ È¿À²À» ÀÌ¿ëÇÑ ÃÖ°í ¼º´ÉÀÇ single cell RNA-seq (ƯÈ÷, RNA ÇÔ·®ÀÌ ¸Å¿ì ³·Àº ¼¼Æ÷ ¹× ÇÙ¿¡ Àû¿ë)
- High library yield - ³ôÀº yieldÀÇ library¸¦ Á¦ÀÛÇÒ ¼ö ÀÖ¾î, NovaSeq Àåºñ¿Í °°Àº high-throughput sequencer¿¡ Àû¿ëÇϰųª, ¿©À¯ºÐÀÇ library´Â ÈÄ¼Ó ½ÇÇè¿¡ ´Ù½Ã ÀÌ¿ëÇÒ ¼ö ÀÖÀ½
- Highly scalable workflow- ´ë·® (High-throughput) ½ÇÇè Àû¿ë°¡´É, ÀÚµ¿È ±â±â Àû¿ë¿¡ ÃÖÀû
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SMART-Seq® mRNA Single Cell Kit¿Í SMART-Seq® mRNA Single Cell LP Kit´Â ³ôÀº ¹Î°¨µµ¿Í ÀçÇö¼ºÀ¸·Î single cell ȤÀº nuclei¸¦ ºÐ¼®ÇÒ ¼ö ÀÖ´Ù. SMART-Seq® mRNA Single Cell LP Kit¸¦ ÀÌ¿ëÇϸé, single cell¿¡¼ RNA¸¦ ÃßÃâÇÏÁö ¾Ê°í, ¹Ù·Î oligo(dT) primer¸¦ ÀÌ¿ëÇØ °íǰÁú full-length cDNA¸¦ Á¦Á¶ÇÑ ÈÄ, lllumina® ±â±â¿¡¼ »ç¿ë °¡´ÉÇÑ library Á¦ÀÛ±îÁö ÁøÇàÇÒ ¼ö ÀÖ´Ù. ÀÌ Á¦Ç°Àº 2 pg total RNA¿Í RNA ÇÔ·®ÀÌ ³·Àº °ÍÀ¸·Î ¾Ë·ÁÁ® ÀÖ´Â ¸»ÃÊÇ÷¾× ´ÜÇÙ¼¼Æ÷ (PBMCs)¿Í °°Àº single cell¿¡µµ ¿ì¼öÇÑ °á°ú¸¦ È®ÀÎÇÏ¿´´Ù.
¶ÇÇÑ single cell ºÐ¼®À» À§ÇØ SMART (Switching Mechanism at 5' End of RNA Template) ±â¼úÀ» ¾÷±×·¹À̵åÇÏ¿© °³¹ßµÈ ½Ã½ºÅÛÀ¸·Î, ´õ ¸¹Àº read¿Í À¯ÀüÀÚ¸¦ °ËÃâÇÒ ¼ö ÀÖ´Ù. SMART-Seq® mRNA Single Cell Kit´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 Single Cell LP Kit¿¡ Æ÷ÇԵǾî ÀÖ´Â library preparation kit´Â enzymatic fragmentation¿Í stem-loop adaptor¸¦ »ç¿ëÇÏ¿©, 2½Ã°£ ³» single tube¿¡¼ purification °úÁ¤ ¾øÀÌ cDNA·ÎºÎÅÍ library¸¦ 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¸ÀÇ ¹æ¹ý ¹× ´ç»ç ±âÁ¸ Á¦Ç° (SMART-Seq® v4)¿¡ ºñÇØ ÀçÇö¼ºÀÌ ³ô°í, ÈξÀ ¿ì¼öÇÒ »Ó ¾Æ´Ï¶ó, ³ôÀº yield·Î Á¦À۵Ǿî high-throughput sequencer¿¡ Àû¿ëÇϰųª, ¿©À¯ºÐÀÇ library¸¦ ÈÄ¼Ó ½ÇÇè¿¡ ´Ù½Ã ÀÌ¿ëÇÒ ¼ö ÀÖ´Ù.

±×¸² 1. SMART-Seq® Single Cell Kit (SSsc)¿Í SMART-Seq2 (homebrew)ÀÇ °á°ú ºñ±³. SMART-Seq2¸¦ ÀÌ¿ëÇßÀ» ¶§ º¸´Ù SMART-Seq® Single Cell KitÀ» ÀÌ¿ëÇßÀ» ¶§, ¹ÌÅäÄܵ帮¾Æ À¯·¡ À¯ÀüÀڷκÎÅÍ mappingµÈ read°¡ ÈξÀ ¸¹¾ÒÀ¸¸ç, º¸´Ù ¸¹Àº À¯ÀüÀÚ°¡ °ËÃâµÊÀ» È®ÀÎÇÏ¿´´Ù.
Lymphoblastoid cell line GM12878ÀÇ single cell·Î SMART-Seq® Single Cell Kit (18 cells) ¹× Smart-seq2 ¹æ¹ý (20 cells)À¸·Î cDNA¸¦ ÇÕ¼º (ÀÌÈÄ 19 cycles PCR ÁõÆø)ÇÑ ÈÄ RNA-Seq library¸¦ Á¦ÀÛÇÏ¿© ½ÃÄö½º¸¦ ºÐ¼®ÇÏ¿´´Ù (¸ðµç »ùÇÃÀº 175¸¸ paired-end reads·Î normalization).
- ±×¸² A : cDNAÀÇ ÇÕ¼º ¹æ¹ý¿¡ µû¶ó mapping °á°ú°¡ ´Ù¸£°í, SMART-Seq2¿¡¼´Â ¹ÌÅäÄܵ帮¾Æ DNA¿¡ ´ëÇÑ mapping ºñÀ²ÀÌ ³ô¾Æ À¯ÀüÀÚ °ËÃâ¿¡ »ç¿ëÇÒ ¼ö ÀÖ´Â read ¼ö°¡ Àû¾ú´Ù.
- ±×¸² B : SMART-Seq® Single Cell kit¿¡¼´Â SMART-Seq2¿¡ ºñÇØ ¸¹Àº À¯ÀüÀÚ°¡ °ËÃâµÇ¾ú´Ù.

±×¸² 2. SMART-Seq® Single Cell Kit (SSsc)¿Í ´ç»ç ±âÁ¸ Á¦Ç°ÀÎ SMART-Seq® v4 (SSv4)¿ÍÀÇ ºñ±³. ´ç»ç ±âÁ¸ Á¦Ç°ÀÎ SMART-Seq® v4À» ÀÌ¿ëÇßÀ» ¶§º¸´Ù SMART-Seq® Single Cell KitÀ» ÀÌ¿ëÇßÀ» ¶§, ´õ ³ôÀº cDNA yield¸¦ ¾ò¾ú°í, read¿¡ °ü°è¾øÀÌ °ËÃâµÇ´Â À¯ÀüÀÚ ¼ö°¡ 60% ÀÌ»óÀÓÀ» È®ÀÎÇÏ¿´´Ù.
- ±×¸² A : Lymphoblastoid cell line (GM22601)¿¡¼ 12°³ÀÇ single cellÀ» ºÐ¸®ÇÏ¿© SMART-Seq® Single Cell Kit°ú SMART-Seq® v4 (Code 634888 ¿Ü)À» »ç¿ëÇÏ¿© cDNA¸¦ ÇÕ¼ºÇÏ¿´´Ù (ÀÌÈÄ 19 cycles PCR ÁõÆø). cDNA yield´Â SMART-Seq® Single Cell Kit¿¡¼ ÈξÀ ³ô¾Ò´Ù.
- ±×¸² B : ÇϳªÀÇ donor·ÎºÎÅÍ ¹ÞÀº PBMC single cell 50°³¸¦ SMART-Seq® v4 ¶Ç´Â SMART-Seq® Single Cell Kit·Î cDNA¸¦ ÇÕ¼ºÇϰí, RNA-Seq library¸¦ Á¦ÀÛÇÏ¿© ½ÃÄö½º ºÐ¼®À» ¼öÇàÇÏ¿´´Ù. ºÐ¼®¿¡ »ç¿ëÇÑ read ¼ö¿¡ °ü°è¾øÀÌ SMART-Seq® Single Cell Kit¿¡¼ SMART-Seq® v4º¸´Ù 1.6¹è ÀÌ»óÀÇ À¯ÀüÀÚ°¡ °ËÃâµÇ¾ú´Ù. SMART-Seq ±â¼úÀ» single cell ºÐ¼®¿ëÀ¸·Î °³¹ßÇÑ SMART-Seq® mRNA Single Cell kitÀÇ °¨µµ°¡ ¶Ù¾î³ª´Ù´Â °ÍÀ» È®ÀÎÇÒ ¼ö ÀÖ¾ú´Ù.

±×¸² 3. SMART-Seq® Single Cell Kit PLUS (SMART-Seq® mRNA Single Cell LPÀÇ ÀÌÀü À̸§)¸¦ ÀÌ¿ëÇÑ library¿¡¼ Nextera® XTº¸´Ù ´õ ³ôÀº yield¿Í À¯ÀüÀÚ °ËÃâ
- ±×¸² A : SMART-Seq® Single Cell Kit¸¦ »ç¿ëÇÏ¿© 10 pgÀÇ ¸¶¿ì½º ³ú total RNA·ÎºÎÅÍ 3¹Ýº¹À¸·Î cDNA¸¦ ÇÕ¼ºÇÏ¿´´Ù.
- ±×¸² B : SMART-Seq® Single Cell PLUS Kit´Â Nextera XTº¸´Ù library yield°¡ ³ô¾Ò´Ù.
- ±×¸² C : SMART-Seq® Single Cell PLUS Kit´Â Nextera XT¿¡ ºñÇØ ´õ ¸¹Àº À¯ÀüÀÚ¸¦ °ËÃâÇÒ ¼ö ÀÖ¾ú´Ù.
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SMART-Seq® Single Cell Kit
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SMART-Seq® Single Cell Kit LP
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634789, 634790, 634785
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634786, 634787, 634788
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±âº» ±¸¼º
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Control Total RNA (1 µg/µl)
10X Lysis Buffer
RNase Inhibitor (40 U/µl)
Nuclease-Free Water
SMART-Seq sc TSO
3′ SMART-Seq CDS Primer II A
SMARTScribe™ II Reverse Transcriptase
SMART-Seq sc First Strand Buffer (5X)
SeqAmp™ DNA Polymerase
SeqAmp CB PCR Buffer (2X)
PCR Primer
Elution Buffer (10 mM Tris-Cl, pH 8.5)
|
LP (Library Prep.)
|
|
10X FE
FE Dilution Buffer
Lib Prep Buffer
Lib Prep Enzyme
Rxn Enhancer
Stem-Loop Adapters
Amplification Buffer
PrimeSTAR® HS DNA Polymerase (5 U/µl)
Nuclease-Free Water
|
* index primer´Â º°µµ·Î ±¸¸Å°¡ ÇÊ¿äÇÕ´Ï´Ù.
>> ȣȯµÇ´Â Unique Dual Index kits
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