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Home > 전제품보기 > NGS 관련 > Ultra Low/Single cell-Seq > SMARTer Ultra Low Input RNA Kit for Sequencing - v3

SMARTer Ultra Low Input RNA Kit for Sequencing - v3

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제조사 제품코드 제품명 용량 가격
(부가세별도)
비고 사용자매뉴얼
Clontech
634850
SMARTer Ultra Low Input RNA Kit for Sequencing - v3
관련학술 관심상품등록 구매하기 라이선스 
48회
7,602,000원  제조사 페이지로 바로가기 93858
Clontech
634851
SMARTer Ultra Low Input RNA Kit for Sequencing - v3
관련학술 관심상품등록 구매하기 라이선스 
96회
12,193,000원  제조사 페이지로 바로가기

"SMART-Seq Ultra Low Input: Single cell sequencing (단일세포 염기서열 분석) 논문리스트 보기"

* 본 제품으로 합성한 cDNA는 Illumina와 Ion Torrent sequencing platform에 모두 적용 가능합니다.

SMARTer Ultra Low Input RNA Kit for Sequencing - v3는 매우 소량의 total RNA (10 pg-10 ng)나 single cell (1-1,000개의 cells)로부터 바로 RNA-Seq (Transcriptome analysis)에 이용할 수 있는 cDNA를 제작할 수 있는 제품이다. 이전 버전의 SMARTer Ultra Low RNA Kit for Illumina Sequencing-HV 제품에 비하여 SMARTer Ultra Low Input RNA Kit for Sequencing - v3는 보다 많은 유전자를 동정할 수 있고, 높은 transcript mapping율을 보일 뿐만 아니라 더 높은 exon 커버율을 보인다 (greater number of genes, higher transcript mapping, and higher exon coverage). 또한 본 제품은 Illumina와 Ion Torrent 등 다양한 회사의 library preparation kit 또는 sequencing 기기에 적용 가능하다.

본 제품의 핵심 기술인 SMART (Switching Mechanism at 5’ End of RNA Template) 법은 세포의 splice junction과 alteranative splicing event를 포함한 전반적인 전사체 연구 (full transcriptome analysis)에 있어 매우 유용한 기술이다. 또한, 본 제품은 high quality RNA (RIN > 8)나 intact cell로부터 특이적으로 mRNA를 증폭할 수 있다.
특징
  • Sample prep made easy - The single-tube protocol works directly on whole cells and preserves sample integrity
  • Unparalleled sensitivity - Start with as little as 1 cell or 10 pg of total RNA (input range: 1-1,000 cells or 10 pg-10 ng of total RNA)
  • Improved cDNA amplification - The SeqAmp DNA Polymerase included in the kit better amplifies high GC-content genes
  • Suitable for multiple sequencing platforms - cDNA libraries are compatible with either Illumina® or Ion Torrent NGS platforms
  • High-quality RNA-seq data - Get a higher number of genes identified, full length gene analysis, typically less than 2% rRNA reads, and improved representation of GC-rich genes


그림 1. Comparison of sequencing metrics generated with either the SMARTer Ultra Low Input RNA for Illumina Sequencing kit (UL-HV) or the SMARTer Ultra Low Input RNA Kit for Sequencing - v3 (UL-v3). cDNA libraries were generated from either 100 pg Human Brain Total RNA (HBR) or 10 pg Mouse Brain Total RNA (MBR) and were sequenced on an Illumina MiSeq platform. The increased sensitivity gained using the UL-v3 protocol is most significant at the lowest RNA inputs.



그림 2. Individual cells or 1,000 cells (HeLa or Jurkat; Panel A and Panel B, respectively) were used as input for the SMARTer Ultra Low Input RNA Kit for Sequencing - v3. The cDNA samples were analyzed on an Agilent 2100 Bioanalyzer. The single main peak indicates the purity and yield of the cDNA (the additional peak at ~1 kb in the HeLa cell samples is the contribution of a single highly expressed gene, FTH1).



그림 3. Sequencing results for libraries made from 100 pg Human Brain Total RNA using either the SMARTer Ultra Low Input RNA for Illumina Sequencing kit (UL-HV) or the SMARTer Ultra Low Input RNA Kit for Sequencing - v3 (UL-v3). Genes were binned by GC content and correlation plots were used to evaluate the two kits. The average gene counts were very reproducible for replicate samples using UL-HV (Panel A) or UL-v3 (Panel B). When the two protocols were compared, genes with a high GC content (red) showed higher expression with the UL-v3 protocol while genes with a median or low GC content (gray and blue, respectively) showed similar expression levels with both protocols (Panel C).



그림 4. The gene body coverage of the cDNA libraries made from 1 or 1,000 HeLa cells (blue and red lines, respectively), or 1 or 1,000 Jurkat cells (green and purple lines, respectively) using the SMARTer Ultra Low Input RNA Kit for Sequencing - v3 was determined using RSeQC. Read coverage was normalized using Excel.



그림 5. cDNA libraries were made from 1 or 1,000 cells (HeLa or Jurkat) using the SMARTer Ultra Low Input RNA Kit for Sequencing - v3 (UL-v3). Results of Illumina library preparation and sequencing. Table IIB. Results of Ion Torrent library preparation and sequencing.
적용
  • Transcriptome sequencing (RNA-Seq)을 위한 소량의 cell 또는 total RNA로부터 cDNA 합성
  • 본 제품으로 합성한 cDNA는 Illumina sequencing platfomr에 적용하기 위하여, Clontech의 Low Input Library Prep Kit (code 634947) 또는 Nextra sample preparation kit을 이용할 수 있다. 또한 Ion Torrent를 이용하기 위해서는 Ion Xpress Plus Fragment Library Kit을 사용 가능하다.
구성품 (자세한 내용은 CoA를 참조하세요)
* SMARTer Ultra Low Input RNA Kit for Sequencing - v3 Components (Cat. No. 634854; not sold separately)
* SeqAmp DNA Polymerase (Cat. No. 638504)

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