주문정보
- - 재고수량은 변동될 수 있습니다.
- - 재고 확인 시 "갱신" 버튼을 누르시면 실시간 재고를 확인하실 수 있습니다.
- - 가격이 ‘별도문의‘ 시, 상단 ‘견적신청’ 버튼을 눌러 문의해주시면 빠른 답변을 받으실 수 있습니다.
| 선택 | Cat.No. | 제품명 | 가격(VAT별도) | 수량 |
|---|
제품특징
□ 특징
● Complete V(D)J sequence information - TCR mRNA의 가변영역(variable regions)의 전체 서열(Full length) 분석
● TCR-alpha, TCR-beta chain을 동시에 또는 개별적으로 NGS Library 제작 가능
● No multiplex PCR required -1회 반응당 단일 프라이머로 각각의 TCR Subunit을 증폭
● Illumina®-ready sequencing libraries - ligation-independent 방식으로 illumina® adaptor 및 index sequence 부착 가능 (Incorporate Illumina® adapter and index sequences in a ligation-independent manner, and multiplex up to 96 libraries in a single flow-cell lane)
□ 제품설명
SMARTer®Mouse TCR a/b Profiling Kit은 T-cell receptor (TCR) repertoire의 NGS 분석을위한 Illumina platform NGS Library 제작 키트이다. 5’-RACE 기술을 접목하여 TCR 전사체의 V(D)J 가변영역(variable region)을 분석하며, mouse spleen, thymus, or PBMCs로부터얻은 10ng-500ng의 total RNA 또는 1,000개-10,000개의purified T-cell을 적용 가능하다. 본 제품을 이용하면 TCR-α, TCR-β chain을 동시에 또는 각 chain에 대한라이브러리를 개별적으로 제작하여 분석할 수 있다. Multiplexing PCR을 이용하는 기존의 TCR profiling 방법과 달리,library amplification의 각 반응은 single primer pair를이용하므로 primer-dimer 생성 가능성을 낮출 수 있다.Multiplex PCR 방식은 서열특이적 편향성을 나타낼 가능성이 있으나 RACE 방식을이용할 경우 이러한 편향성의 문제를 덜 수 있다. 본 제품은 LNA 기술이 결합된 SMART (Switching Mechanism at 5’ End of RNA Template)법을 이용하므로 민감도가 매우 높고 편향성이 적은 TCR mRNA sequence 분석을 할 수 있다.
그림 1. Library preparation workflow andPCR strategy for TCR profiling using the SMARTer approach.
그림 2. Assessment of TCR diversity inmouse spleen samples with the SMARTer Mouse TCR a/b Profiling Kit. Panel A. To compare TCR diversity betweenthe spleen"s general T-cell population and purified CD4+ T cells, totalsplenocytes were isolated from spleen and divided into two groups. The firstgroup was left untouched, and the second group was subject to CD4+ T-cellpurification (see Methods section for details). Total RNA was extracted fromthe total splenocyte group and the group enriched for CD4+ T cells, and eachRNA sample was used for library preparation. Panel B. Proportion of on-targetreads mapping to TCR-α or TCR-β CDR3 sequences. Panel C. Rarefaction curvesshowing the relationship between read depth and the number of unique clonotypesidentified for TCR-α (left) and TCR-β (right). When a curve starts to flattenoff, the number of unique clonotypes identified is reaching saturation. Dashedlines show the theoretical number of clonotypes as the number of readsincreases.
그림 3. Assessmentof TCR-α clonotype diversity in various tissues of transgenic mice of differentages with the SMARTer Mouse TCR a/b Profiling Kit. Panel A. Thepercentage of reads mapping to TCR-α CDR3 sequences identified in the thymus,spleen, peripheral lymph nodes (PLN), mesenteric lymph nodes (MLN), and largeintestine (LI) is consistent across the 16-week-old, 34-week-old, and38-week-old mice. Panel B. The immune tissues showed the highest TCR-αclonotypic diversity, while the large intestine showed the lowest diversity.This indicates that the large intestine has the lowest T-cell infiltrate. PanelC. Abundance of the TRAV3-2-TRAJ58 clonotype, expressed as a percentage of allreads mapping to any TCR-α or TCR-β clonotype, across tissue types and ages.Panel D. Abundance of the TRAV16N-TRAJ56 clonotype, expressed as a percentageof all reads mapping to any TCR-α or TCR-β clonotype, across tissue types inthe 38-week-old mouse.
