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Home > ÀüÁ¦Ç°º¸±â > NGS °ü·Ã > Immune-Seq > Human scTCR a/b Profiling Kit
´ÜÀÏ T ¼¼Æ÷ (Single T cell)·ÎºÎÅÍ TCR ¥á¥â pairing ºÐ¼®

Human scTCR a/b Profiling Kit

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Á¦Á¶»ç Á¦Ç°ÄÚµå Á¦Ç°¸í ¿ë·® °¡°Ý
(ºÎ°¡¼¼º°µµ)
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Clontech
634431
SMARTer¢ç Human scTCR a/b Profiling Kit
°ü·ÃÇмú ±¸¸ÅÇϱ⠶óÀ̼±½º 
96ȸ
7,494,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â x100088, x97970
Clontech
634432
SMARTer¢ç Human scTCR a/b Profiling Kit
°ü·ÃÇмú ±¸¸ÅÇϱ⠶óÀ̼±½º 
480 ȸ
23,191,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â

  • Flexible workflow: FACS ¶Ç´Â ¸Å´º¾ó ¹æ½ÄÀ¸·Î ºÐ¸®ÇÑ ´ÜÀϼ¼Æ÷ (single cell)·ÎºÎÅÍ Illumina¢ç NGS Library Á¦ÀÛ (MiSeq¢ç ±ÇÀå)
  • Ease of use: 12°³ÀÇ library¿¡ 96°³ÀÇ ¼¼Æ÷¸¦ pooling °¡´ÉÇϹǷÎ, single flow-cell lane¿¡¼­ ºÐ¼® °¡´É
  • Sensitivity: SMART ±â¼ú°ú RACE ¹æ½ÄÀ» Á¶ÇÕÇÏ¿© low-abundance TCR º¯ÀÌü °ËÃâ
  • Specificity: Human TCR¥á, TCR¥â chainÀÇ V(D)J regionÀÇ ¼­¿­°ú Á¤È®ÇÑ pairing Á¤º¸ ȹµæ °¡´É
Á¦Ç°¼³¸í
¿©·¯ °³ÀÇ T-cell »ùÇà (Bulk T-cell sample)À» Çѹø¿¡ ºÐ¼®ÇÏ´Â °ÍÀº T-cell receptor (TCR) repertoire ´Ù¾ç¼ºÀ» ºÐ¼®ÇÏ´Â µ¥¿¡ µµ¿òÀÌ µÇÁö¸¸ T-cellÀÇ Æ¯Á¤ alpha-beta receptor chain paringÀ» È®ÀÎÇÒ ¼ö´Â ¾ø´Ù. TCRÀÇ Æ¯ÀÌÀûÀÎ alpha-beta paringÀº Ç׿øÆ¯À̼º°ú ¿¬°üÀÌ Àֱ⠶§¹®¿¡ pairing Á¤º¸¸¦ È®ÀÎÇÏ´Â °ÍÀº Ç׿øÀÎ½Ä (antigen recognition)ºÐ¼®, Ç¥Àû¸é¿ª¿ä¹ý (targeted immunotherapy)ÀÇ È¿À²ÀûÀÎ TCR µðÀÚÀλӸ¸ ¾Æ´Ï¶ó T-cell populationÀÇ Á¶»ó°ü°è¸¦ ±Ô¸íÇϱâ À§ÇÏ¿© ¸Å¿ì Áß¿äÇÏ´Ù. ±âÁ¸ÀÇ ´ÜÀÏ T-cell (single T-cell)À» ÀÌ¿ëÇϸé,
alpha-beta pairingÀ» È®ÀÎÇÒ ¼ö´Â ÀÖÀ¸³ª, ´ë·®ÀÇ ¸¹Àº ¼¼Æ÷¸¦ ºÐ¼®ÇØ¾ß Çϱ⠶§¹®¿¡ »ó´çÈ÷ ºñ¿ëÀÌ ³ô°í º¹ÀâÇÑ ºÐ¼® °úÁ¤ÀÌ ÇÊ¿äÇß´Ù.
SMARTer¢ç Human scTCR a/b Profiling Kit (Code 634431)Àº SMART¢â cDNA ÇÕ¼º±â¼ú°ú 8Á¾·ùÀÇ unique indexed oligo¸¦ »ç¿ëÇÏ¿© manual ¹æ½Ä ¶Ç´Â FACS µîÀ» ÀÌ¿ëÇÏ¿© ºÐ¸®ÇÑ
96°³ÀÇ T-cellÀ» 12°³ÀÇ NGS sequencing library·Î Á¦ÀÛÇÒ ¼ö ÀÖ¾î ±âÁ¸ Single T-cell ºÐ¼®¹æ¹ýÀÇ ¹®Á¦Á¡À» ±Øº¹ÇÏ¿´´Ù. ¶ÇÇÑ, 96°³ »ùÇÃÀ» À§ÇÑ MultiplexingÀÌ °¡´ÉÇϸç ÀÌ´Â ÇϳªÀÇ flow cell-lane¿¡¼­ ºÐ¼® °¡´ÉÇÏ´Ù.
SMARTer¢ç Human TCR a/b Profiling Kit (Code 635014) Á¦Ç°°ú ¸¶Âù°¡Áö·Î, º» Á¦Ç°Àº SMART (Switching Mechanism at 5¡¯ End of RNA Template) cDNA ÇÕ¼º¹ý°ú
RACE (Rapid Amplification of cDNA Ends) ¹æ½ÄÀ» ÀÌ¿ëÇϹǷÎ, TCR-¥á, ¥â chainÀÇ °¡º¯¿µ¿ª (variable region (V(D)J))À» ƯÀÌÀûÀ¸·Î ¼±ÅÃÇÏ¿© Illumina¢ç NGS¿ë Library¸¦ Á¦ÀÛÇÒ ¼ö ÀÖ´Ù.

* º» Á¦Ç°À» ÀÌ¿ëÇÏ¿© ½ÃÄö½Ì µ¥ÀÌÅÍ ºÐ¼® ½Ã¿¡´Â SMARTer¢â Human scTCR Demultiplexer À¥ÆäÀÌÁö¸¦ ÂüÁ¶ÇϽʽÿÀ.


±×¸² 1. SMARTer¢ç Human scTCR a/b Profiling Kit workflow and pooling strategy
Panel A. SMARTer¢ç Human scTCR a/b Profiling Kit workflow
Panel B. SMART-Seq Indexed Oligo Master Mix Ç÷¹ÀÌÆ® »óÀÇ ¹èÄ¡. Samples are pooled by column, such that each pool contains eight cells each with a differently indexed SMART-Seq Indexed Oligo. Different combinations of the Forward and Reverse HT indexes are used during PCR 2 to allow multiplexing of the samples into a single flow-cell lane (see the User Manual for more details).


±×¸² 2. Benefits of studying T cells at the single-cell level.
Panel A. alpha chain (TCR-¥á)°ú beta chain (TCR-¥â)À¸·Î ±¸¼ºµÈ T-cell receptorÀÇ ¸ð½Äµµ
Panel B. T-cell Áý´Ü (Bulk T-cells)ÀÇ ¼­¿­ ºÐ¼® µ¥ÀÌÅͷκÎÅÍ alpha-beta paring Á¤º¸¸¦ ¾Ë±â ¾î·Æ´Ù.
Rare clonotypes can allow for assessing the pairing of some cells (clonotypes in orange). However, for the more highly expressed clonotypes (TCRA V2J1, TCRA V1J2, TCRB V3D1J1, TCRB V2D1J2) there are four possible pairing combinations.


±×¸² 3. Analysis of a mixed cell population from a 96-well plate.
Panel A. Cell-type calling based on the identified clonotypes for each well. The seven omitted cells did not have clonotype calls for either TCR-¥á or TCR-¥â with read numbers that were above the threshold.
Panel B. Analysis of pairing information. Paired TCR-¥á¥â clonotypes were obtained for 34 cells in the plate.
Panel C. The alpha, beta, or alpha-beta pairing information represented as a percent of cells analyzed. Omitted cells were not included in this analysis.
Application
  • Human TCR repertoire analysis in single cells (TCR-alpha and TCR-beta subunits)
  • TCR alpha-beta pairing determination in single cells
±¸¼ºÇ° (ÀÚ¼¼ÇÑ ³»¿ëÀº CoA¸¦ ÂüÁ¶Çϼ¼¿ä)
  • SMARTer Human scTCR a/b Profiling Kit Components
  • TCR a/b Human Indexing Primer Set HT for Illumina
º¸Á¸
  • Control Jurkat Total RNA, SMART-Seq Indexed Oligos: - 70¡É
  • 10X Lysis Buffer, Nuclease-Free water: - 20¡É (À¶ÇØ ÈÄ, 4¡É º¸°ü)
  • Elution Buffer: - 20¡É (À¶ÇØ ÈÄ, ½Ç¿Â º¸°ü)
  • ±âŸ: - 20¡É

Keyword : TCR,CDR3,T cell receptor,repertoire,variable region,Immune cell,Immune-seq,Immunology,,smart-seq,SMART,NGS,Next generation sequencing,cancer,Cancer research,¾Ï¿¬±¸,vaccine,¹é½Å,¹é½Å¿¬±¸

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