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TCR/BCR Profiling
> Mouse TCR a/b Profiling Kit
Mouse TCR a/b Profiling Kit
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Clontech
634403
SMARTer¢ç Mouse TCR a/b Profiling Kit
48ȸ
(Package)
5,132,800¿ø
6,416,000¿ø
11.01 ~ 12.27
Clontech
634404
SMARTer¢ç Mouse TCR a/b Profiling Kit
96ȸ
(Package)
8,365,600¿ø
10,457,000¿ø
11.01 ~ 12.27
Product index
Overview
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TCR/BCR Profiling
Mouse TCR a/b Profilin..
Mouse BCR IgG H/K/L Pr..
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Cell culture ÇʼöÇ° - ..
¼¼Æ÷¹è¾ç½Ã Mycoplasma ..
MycoAlert PLUS mycopla..
MycoAlert Mycoplasma D..
MycoZap Antibiotics
MycoZap Mycoplasma Eli..
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Complete V(D)J sequence information -
TCR mRNAÀÇ °¡º¯¿µ¿ª(variable regions)ÀÇ Àüü ¼¿(Full length) ºÐ¼®
TCR-alpha, TCR-beta chain
À» µ¿½Ã¿¡ ¶Ç´Â °³º°ÀûÀ¸·Î NGS Library Á¦ÀÛ °¡´É
No multiplex PCR required -
1ȸ ¹ÝÀÀ´ç ´ÜÀÏ ÇÁ¶óÀ̸ӷΠ°¢°¢ÀÇ TCR SubunitÀ» ÁõÆø
Illumina¢ç-ready sequencing libraries -
ligation-independent ¹æ½ÄÀ¸·Î illumina¢ç adaptor ¹× index sequence ºÎÂø °¡´É (Incorporate Illumina¢ç adapter and index sequences in a ligation-independent manner, and multiplex up to 96 libraries in a single flow-cell lane)
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SMARTer¢ç Mouse TCR a/b Profiling KitÀº T-cell receptor (TCR) repertoireÀÇ NGS ºÐ¼®À» À§ÇÑ Illumina platform NGS Library Á¦ÀÛ Å°Æ®ÀÌ´Ù. 5¡¯-RACE ±â¼úÀ» Á¢¸ñÇÏ¿© TCR Àü»çüÀÇ V(D)J °¡º¯¿µ¿ª (variable region)À» ºÐ¼®Çϸç, mouse spleen, thymus, or PBMCs·ÎºÎÅÍ ¾òÀº 10ng-500ngÀÇ total RNA ¶Ç´Â 1,000°³-10,000°³ÀÇ purified T-cellÀ» Àû¿ë °¡´ÉÇÏ´Ù. º» Á¦Ç°À» ÀÌ¿ëÇϸé TCR-¥á, TCR-¥â chainÀ» µ¿½Ã¿¡ ¶Ç´Â °¢ chain¿¡ ´ëÇÑ ¶óÀ̺귯¸®¸¦ °³º°ÀûÀ¸·Î Á¦ÀÛÇÏ¿© ºÐ¼®ÇÒ ¼ö ÀÖ´Ù.
Multiplexing PCRÀ» ÀÌ¿ëÇÏ´Â ±âÁ¸ÀÇ TCR profiling ¹æ¹ý°ú ´Þ¸®, library amplificationÀÇ °¢ ¹ÝÀÀÀº single primer pair¸¦ ÀÌ¿ëÇϹǷΠprimer-dimer »ý¼º °¡´É¼ºÀ» ³·Ãâ ¼ö ÀÖ´Ù. Multiplex PCR ¹æ½ÄÀº ¼¿Æ¯ÀÌÀû ÆíÇ⼺À» ³ªÅ¸³¾ °¡´É¼ºÀÌ ÀÖÀ¸³ª RACE ¹æ½ÄÀ» ÀÌ¿ëÇÒ °æ¿ì ÀÌ·¯ÇÑ ÆíÇ⼺ÀÇ ¹®Á¦¸¦ ´ú ¼ö ÀÖ´Ù.
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emplate)¹ýÀ» ÀÌ¿ëÇϹǷΠ¹Î°¨µµ°¡ ¸Å¿ì ³ô°í ÆíÇ⼺ÀÌ ÀûÀº TCR mRNA sequence ºÐ¼®À» ÇÒ ¼ö ÀÖ´Ù.
±×¸² 1. Library preparation workflow and PCR strategy for TCR profiling using the SMARTer approach.
±×¸² 2. Assessment of TCR diversity in mouse spleen samples with the SMARTer Mouse TCR a/b Profiling Kit.
Panel A. To compare TCR diversity between the spleen's general T-cell population and purified CD4+ T cells, total splenocytes were isolated from spleen and divided into two groups. The first group was left untouched, and the second group was subject to CD4+ T-cell purification (see Methods section for details). Total RNA was extracted from the total splenocyte group and the group enriched for CD4+ T cells, and each RNA sample was used for library preparation. Panel B. Proportion of on-target reads mapping to TCR-¥á or TCR-¥â CDR3 sequences. Panel C. Rarefaction curves showing the relationship between read depth and the number of unique clonotypes identified for TCR-¥á (left) and TCR-¥â (right). When a curve starts to flatten off, the number of unique clonotypes identified is reaching saturation. Dashed lines show the theoretical number of clonotypes as the number of reads increases.
±×¸² 3. Assessment of TCR-¥á clonotype diversity in various tissues of transgenic mice of different ages with the SMARTer Mouse TCR a/b Profiling Kit.
Panel A. The percentage of reads mapping to TCR-¥á CDR3 sequences identified in the thymus, spleen, peripheral lymph nodes (PLN), mesenteric lymph nodes (MLN), and large intestine (LI) is consistent across the 16-week-old, 34-week-old, and 38-week-old mice. Panel B. The immune tissues showed the highest TCR-¥á clonotypic diversity, while the large intestine showed the lowest diversity. This indicates that the large intestine has the lowest T-cell infiltrate. Panel C. Abundance of the TRAV3-2-TRAJ58 clonotype, expressed as a percentage of all reads mapping to any TCR-¥á or TCR-¥â clonotype, across tissue types and ages. Panel D. Abundance of the TRAV16N-TRAJ56 clonotype, expressed as a percentage of all reads mapping to any TCR-¥á or TCR-¥â clonotype, across tissue types in the 38-week-old mouse.
Applications
Mouse TCR repertoire analysis (TCR-alpha and TCR-beta subunits)
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SMARTer Mouse TCR a/b Profiling Kit Components
TCR a/b Mouse Indexing Primer Set HT for Illumina - 12
Keyword :
TCR/BCR Profiling
Mouse TCR a/b Profiling Kit
Mouse BCR IgG H/K/L Profiling Kit