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Home > ÀüÁ¦Ç°º¸±â > NGS °ü·Ã > Immune-Seq > [New]UMI Àû¿ë Mouse TCR profiling(v2)

[New]UMI Àû¿ë Mouse TCR profiling(v2)

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Á¦Á¶»ç Á¦Ç°ÄÚµå Á¦Ç°¸í ¿ë·® °¡°Ý
(ºÎ°¡¼¼º°µµ)
ºñ°í »ç¿ëÀڸŴº¾ó
Clontech
634815
SMART-Seq¢ç Mouse TCR (with UMIs)
°ü·ÃÇмú ±¸¸ÅÇϱ⠶óÀ̼±½º 
24ȸ
(Package)
2,705,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â x346022
Clontech
634816
SMART-Seq¢ç Mouse TCR (with UMIs)
°ü·ÃÇмú ±¸¸ÅÇϱ⠶óÀ̼±½º 
96ȸ
(Package)
9,028,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â
Clontech
634814
SMART-Seq¢ç Mouse TCR (with UMIs)
°ü·ÃÇмú ±¸¸ÅÇϱ⠶óÀ̼±½º 
384ȸ (634816 x 4)
32,501,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â

¡Ø º¯°æ»çÇ×

º» Á¦Ç°Àº ±âÁ¸ SMARTer¢ç Mouse TCR a/b Profiling KitÀÇ ¾÷±×·¹À̵å Á¦Ç°ÀÔ´Ï´Ù.
±âÁ¸ Á¦Ç°º¸´Ù input range°¡ ³Ð¾îÁ® ´Ù¾çÇÑ »ùÇÿ¡ Àû¿ë °¡´ÉÇØÁ³À¸¸ç, UMI (unique molecular identifiers)°¡ Ãß°¡µÇ¾ú½À´Ï´Ù.

Á¦Ç°»ç¿ë À¯¿¬¼ºÀ» À§ÇØ Á¦Ç° ±¸¼º¿¡¼­ index°¡ Á¦¿ÜµÇ¾ú½À´Ï´Ù. ÇÊ¿ä½Ã º°µµ±¸¸Å ¹Ù¶ø´Ï´Ù.
>> ȣȯµÇ´Â Unique Dual Index kits

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

  • ³ÐÀº ¹üÀ§ÀÇ »ùÇà input Àû¿ë: 1 ng-1 §¶ÀÇ total RNA (mouse spleen, T cell), 10 ng-1 §¶ÀÇ total RNA (mouse whole blood, bone marrow, thymus), 100-10,000°³ÀÇ purified mouse T cell
  • ³ôÀº °¨µµ¿Í ½ÉÇÃÇÑ ¿öÅ©Ç÷οì: TCR ¥á ¿Í TCR ¥â subunitÀÇ library¸¦ Á¦ÀÛÇϴµ¥ µ¿ÀÏÇÑ primer¸¦ ÀÌ¿ëÇϱ⠶§¹®¿¡ µ¿½Ã¿¡ °¢°¢ÀÇ TCR clonotypeÀ» ¹Î°¨ÇÏ°Ô °ËÃâÇÒ ¼ö ÀÖÀ½
  • Sequencing flexibility: full-length V(D)J ¶Ç´Â CDR3¸¸ ¼±ÅÃÇؼ­ sequencing°¡´É
Á¦Ç°¼³¸í
SMART-Seq¢ç Mouse TCR (with UMIs)´Â T-cell receptor (TCR) repertoire NGS ºÐ¼®À» À§ÇÏ¿© Illumina platform NGS Library¸¦ Á¦ÀÛÇÒ ¼ö ÀÖ´Â Á¦Ç°ÀÌ´Ù. SMART (Switching Mechanism at 5¡¯ end of RNA Template) ±â¼úÀ» Àû¿ëÇÏ¿© ³ôÀº °¨µµ·Î bias ¾øÀÌ TCR ¼­¿­À» °ËÃâÇÒ ¼ö ÀÖÀ¸¸ç, 5¡¯-RACE Á¢±Ù¹æ½ÄÀ» °áÇÕÇÏ¿© ¸¶¿ì½º TRA ¹× TRB À¯ÀüÀÚÀÇ full-length V(D)J °¡º¯¿µ¿ªÀ» ĸóÇÒ ¼ö ÀÖ´Ù.
º» Á¦Ç°Àº ´Ù¾çÇÑ RNA input (RIN>7)¿¡ Àû¿ëÇÒ ¼ö ÀÖÀ¸¸ç, mouse spleen ¶Ç´Â T cell¿¡¼­ ¾òÀº 1 ng-1 §¶ÀÇ total RNA, mouse whole blood, bone marrow, thymus¿¡¼­ ¾òÀº 10 ng-1 §¶ÀÇ total RNA, ±×¸®°í 100-10,000°³ÀÇ Á¤Á¦µÈ mouse T cell·ÎºÎÅÍ °íÇ°ÁúÀÇ sequencing library¸¦ ÁغñÇÒ ¼ö ÀÖ°í À̸¦ ÅëÇØ ¥á, ¥â chain °¢°¢ÀÇ ´Ù¾ç¼º Á¤º¸¸¦ ¾òÀ» ¼ö ÀÖ´Ù.
ÀÌ profiling kit´Â UMI (unique molecular identifiers)¸¦ Æ÷ÇÔÇÏ¿© PCR duplicates, sequencing error¿¡ ÀÇÇØ ¹ß»ýµÈ read¸¦ Á¦°ÅÇÒ ¼ö Àֱ⠶§¹®¿¡ º¸´Ù Á¤È®ÇÏ°í ½Å·Ú¼º ÀÖ´Â °á°ú¸¦ ¾òÀ» ¼ö ÀÖ´Ù. UDI (Unique Dual Index kits) (Code 634752-634756, º°µµ±¸¸Å)¸¦ »ç¿ëÇÏ¿© ÃÖ´ë 384°³ÀÇ multiplexed Illumina library¸¦ »ý¼ºÇÒ ¼ö ÀÖ´Ù.


±×¸² 1. ´Ù¾çÇÑ »ùÇà ŸÀÔ¿¡¼­ mTCRv2 kit (SMART-Seq¢ç Mouse TCR (with UMIs)) ¼º´É Æò°¡
A. mTCRv2 ¿öÅ©Ç÷ο츦 »ç¿ëÇÏ¿© 10, 100, 250, 500, ±×¸®°í 1,000 ngÀÇ mouse spleen RNA·Î TCR library¸¦ ÁغñÇßÀ¸¸ç, TCR ¥á (ÆĶû), TCR ¥â (º¸¶ó) chainÀ» ºÐ¼®ÇÏ¿´´Ù. TCR library´Â Illumina NextSeqÀ¸·Î 2 x 150 bp read¸¦ »ý¼ºÇÏ¿´´Ù (sequencing output: ~1,500¸¸ read).
B. ¼­·Î ´Ù¸¥ »ùÇà ŸÀÔ¿¡¼­ ºÐ¸®µÈ 4Á¾·ùÀÇ 200 ng RNA¸¦ »ç¿ëÇÏ¿© mTCRv2 ¿öÅ©Ç÷ο츦 ¼öÇàÇÏ¿´´Ù. TCR library´Â NextSeqÀ¸·Î sequencingÇÏ¿© 2 x 150 bp read¸¦ »ý¼ºÇÏ¿´´Ù (sequencing output: ~2,000¸¸ read). µ¥ÀÌÅÍ´Â Cogent NGS Immune Profiler software¸¦ »ç¿ëÇÏ¿© ºÐ¼®ÇÏ¿´À¸¸ç, °¢ °³º° input amount ¶Ç´Â »ùÇà ŸÀÔ¿¡ ´ëÇÑ clonotype ¼ö¸¦ ¸·´ë±×·¡ÇÁ·Î ³ªÅ¸³Â´Ù (TRA = TCR ¥á chain, TRB = TCR ¥â chain).
±âÁ¸ Á¦Ç°°ú Â÷ÀÌÁ¡

Á¦Ç°

SMARTer¢ç Mouse TCR a/b Profiling Kit (mTCRv1)

SMART-Seq¢ç Mouse TCR (with UMIs) (mTCRv2)

Code

634402, 634403, 634404

634815, 634816, 634814

Input

¤ý10 ng - 500 ng RNA (spleen, thymus, PBMC)
1,000 - 10,000 purified T cells

¤ý 1 ng - 1 §¶ RNA (spleen, T cells)
¤ý 10 ng - 1 §¶ RNA (whole blood, bone marrow, thymus)
¤ý 100 - 10,000 purified T cells

UMIs

¹ÌÀû¿ë

Àû¿ë

UDIs

ȣȯµÇÁö ¾ÊÀ½

ȣȯµÊ (º°µµ±¸¸Å)

Sequencing
À¯¿¬¼º

2 x 300 bpÀ¸·Î¸¸ Sequencing °¡´É

Full-length V(D)J ¶Ç´Â CDR3¸¸ Sequencing °¡´É

Bioinformatics tool Á¦°ø

´ç»ç ¹«·á tool Á¦°ø
´ç»ç ¹«·á Cogent Immune Viewer* Á¦°ø

Àû¿ë
Mouse TCR repertoire analysis (TCR ¥á and TCR ¥â subunits) ¿¬±¸ºÐ¾ß
¤ý ´Ù¾çÇÑ °¨¿°¼º Áúȯ ¿¹¹æ ¶Ç´Â °³¼±¿¡ °ü·ÃµÈ TCR ¹ÙÀÌ¿À¸¶Ä¿ ½Äº°À» À§ÇÑ º´¿øü ƯÀÌÀû TCR ºÐ¼®
¤ý T¸²ÇÁ±¸ÀÇ À̵¿, À¯Áö ¹× Ç׻󼺿¡ ±âÃÊ°¡ µÇ´Â Á¶Á÷°ú ¼øȯ°è¿¡¼­ TCR Ŭ·Ð ±¸¼º ¹× ´Ù¾ç¼º ¿¬±¸
¤ý Immune aging dynamics¿¡ ´ëÇÑ longitudinal analysis, ¶Ç´Â TCR repertoires º¯È­°¡ ³ëÀΠȯÀÚÀÇ ¹é½Å ¹× ±âŸ Ä¡·á¹ÝÀÀ¿¡ ¾î¶² ¿µÇâÀ» ÁÙ ¼ö ÀÖ´ÂÁö ¿¬±¸
±¸¼ºÇ° (ÀÚ¼¼ÇÑ ³»¿ëÀº CoA¸¦ ÂüÁ¶Çϼ¼¿ä)
º¸°ü Á¶°Ç ¹× Á¦Ç° ±¸¼ºÇ°, ±â¼ú »ç¾ç¿¡ ´ëÇÑ ÀÚ¼¼ÇÑ ³»¿ëÀº Á¦Ç°ÀÇ Certificate of Analysis¸¦ ÂüÁ¶Çϼ¼¿ä. Å°Æ® ±¸¼ºÇ°À» È®ÀÎÇÏ·Á¸é Kit Components List¸¦ ÂüÁ¶Çϼ¼¿ä.
- CoA¿Í MSDS¸¦ È®ÀÎÇÏ´Â ¹æ¹ý (Ŭ¸¯)

Keyword :

Immune-Seq
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