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Home > ÀüÁ¦Ç°º¸±â > NGS °ü·Ã > Stranded RNA-seq > ÀÓ»ó»ùÇà ÃÖÀû Pico Input Mammalian(v2)
Picogram RNA·ÎºÎÅÍ Strand-specific illumina NGS library Á¦ÀÛ (version update)

ÀÓ»ó»ùÇÃ ÃÖÀû Pico Input Mammalian(v2)

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Clontech
634411
SMARTer¢ç Stranded Total RNA-Seq Kit v2 - Pico Input Mammalian
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Clontech
634412
SMARTer¢ç Stranded Total RNA-Seq Kit v2 - Pico Input Mammalian
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Clontech
634413
SMARTer¢ç Stranded Total RNA-Seq Kit v2 - Pico Input Mammalian
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9,200,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â
Clontech
634414
SMARTer¢ç Stranded Total RNA-Seq Kit v2 - Pico Input Mammalian
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º» Á¦Ç°Àº SMART-Seq¢ç Total RNA Pico Input (ZapR¢â Mammalian) (Code 634357, 634358, 634359) Á¦Ç°À¸·Î ¸®´º¾óµÇ¾ú½À´Ï´Ù.
ÀÚ¼¼ÇÑ ³»¿ëÀº ´ÙÄ«¶óÄÚ¸®¾Æ (02-2081-2510)·Î ¹®ÀÇ ÁÖ¼¼¿ä.

* º» Á¦Ç°Àº SMARTer¢ç Stranded Total RNA-Seq Kit - Pico Input MammalianÀÇ ¾÷±×·¹À̵å Á¦Ç°À̸ç,
SMARTer¢ç Stranded Total RNA-Seq Kit v3 - Pico Input MammalianÀ¸·Î ¾÷±×·¹ÀÌµå µÇ¾ú½À´Ï´Ù.
ÀÚ¼¼ÇÑ ³»¿ëÀº ´ÙÄ«¶óÄÚ¸®¾Æ °í°´Áö¿ø (TEL. 02-2081-2510 / mail. support@takara.co.kr)À¸·Î ¿¬¶ô ¹Ù¶ø´Ï´Ù.

  • Picogram ¼öÁØÀÇ ±Ø¼Ò·® »ùÇà Àû¿ë: 250pg - 10ng total RNA (Human, mouse, rat)
  • FFPE, LCM »ùÇÿ¡¼­ ÃßÃâÇÑ RNA ¶Ç´Â Cell-free RNA µîÀÇ ºÐÇØµÈ RNA »ùÇà Àû¿ë °¡´É
  • Improved sequencing performance: PhiX controlÀ» Ãß°¡ÇÏÁö ¾Ê¾Æµµ ³ôÀº ¼öÁØÀÇ cluster passing filter (%PF)¸¦ ¾òÀ» ¼ö ÀÖÀ½
  • Fast, streamlined protocol: 6½Ã°£ À̳» Illumina¢ç NGS Library Á¦ÀÛ °¡´É
  • PCR ¹öÆÛÀÇ Á¶¼ºÀ» º¯ÇüÇÏ¿© Library Á¤Á¦ È¿À² Çâ»ó
Á¦Ç°¼³¸í
SMARTer¢ç Stranded Total RNA-Seq Kit v2 - Pico Input Mammalian Àº 250pg-10ng ¼öÁØÀÇ Human, mouse, rat total RNA·ÎºÎÅÍ Strand-specific Illumina¢ç NGS library¸¦ È¿À²ÀûÀ¸·Î Á¦ÀÛÇÒ ¼ö ÀÖ´Ù. º» Á¦Ç°Àº ±Ø¹Ì·®ÀÇ total RNA¸¦ ÀÌ¿ëÇÒ ¼ö ÀÖÀ» »Ó¸¸ ¾Æ´Ï¶ó FFPE, LCM »ùÇðú °°ÀÌ RNA integrity°¡ ÇöÀúÈ÷ ³·Àº »ùÇÿ¡¼­µµ NGS Library¸¦ Á¦ÀÛ °¡´ÉÇÏ´Ù. ±âÁ¸ÀÇ Version 1 Á¦Ç°¿¡ ºñÇÏ¿© Version 2 Á¦Ç°Àº Illumina¢ç SequencingÀÇ È¿À²À» º¸´Ù Çâ»ó½ÃÄ×À¸¸ç, ¿ªÀü»ç ¹ÝÀÀ°ú PCR ÁõÆø °úÁ¤¿¡¼­ Illumina¢ç index¿Í adaptor¸¦ ºÎ°¡ÇÏ¿© 6½Ã°£ À̳»¿¡ strand-specific RNA-Seq library¸¦ Á¦ÀÛ °¡´ÉÇÏ´Ù.

Ribosomal RNA (rRNA)´Â total RNAÀÇ ¾à 90%¸¦ Â÷ÁöÇÏ´Â RNA·Î½á NGS library Á¦ÀÛ Àü, rRNA¸¦ Á¦°Å(depletion)ÇÔÀ¸·Î½á Á¦ÀÛºñ¿ëÀ» Àý°¨ÇÒ ¼ö ÀÖÀ» »Ó¸¸ ¾Æ´Ï¶ó mapping statistics ¶ÇÇÑ Çâ»ó½Ãų ¼ö ÀÖ´Ù. ÇÏÁö¸¸ ¸Å¿ì ±Ø¼Ò·®ÀÇ Total RNA¸¦ ÀÌ¿ëÇÒ °æ¿ì ÀϹÝÀûÀÎ ¹æ¹ýÀÇ rRNA depletionÀº NGS library Á¦Á¶ÇÏ´Â µ¥ ÇÊ¿äÇÑ ¾ç¿¡ ¹ÌÄ¡Áö ¸ø ÇÏ°Ô µÈ´Ù. º» Á¦Ç°¿¡ »ç¿ëµÈ ±â¼úÀº mammalian rRNA¿Í mitochondrial RNA ƯÀÌÀûÀÎ Probe¸¦ ÀÌ¿ëÇÏ¿© rRNA derived cDNA¸¦ Á¦°ÅÇÏ¿© º¸´Ù ³ôÀº mapping statistics¸¦ ¾òÀ» ¼ö ÀÖ´Ù.

SMARTer¢ç Stranded Total RNA-Seq Kit v2 - Pico Input MammalianÀº two channel SBS ±â¼úÀ» Àû¿ëÇÑ Illumina¢ç NextSeq, MiniSeq, HiSeq 3000/4000À» ÀÌ¿ëÇÏ´Â °æ¿ì¿¡µµ PhiX Control Spike-in RNAÀÇ Ã·°¡ ¾øÀÌ ³ôÀº ºñÀ²ÀÇ Cluster passing filter (%PF)¸¦ ¾òÀ» ¼ö ÀÖ´Ù. ÀÌ´Â 1ȸ Running´ç, º¸´Ù »ý¹°ÇÐÀûÀ¸·Î ÀǹÌÀÖ´Â sequencing reads¸¦ ¾òÀ» ¼ö ÀÖÀ¸¹Ç·Î ÀüüÀûÀ¸·Î sequencing cost¸¦ ÁÙÀÏ ¼ö ÀÖ´Ù. ¶ÇÇÑ º» Á¦Ç°Àº ±âÁ¸ Á¦Ç°¿¡ ºñÇÏ¿© »õ·Î¿î PCR Buffer¸¦ ÀÌ¿ëÇϹǷΠº¸´Ù ³ôÀº NGS Library Á¤Á¦ È¿À²À» ±â´ëÇÒ ¼ö ÀÖ´Ù.

º» Á¦Ç°Àº 12, 48, 96, 192ȸ Á¦Ç°À¸·Î Á¦°øµÇ¸ç, High-throughput Á¦Ç°ÀÎ 96ȸ¿Í 192ȸ Á¦Ç° ³»¿¡´Â 12°³ÀÇ reverse primer¿Í 8°³ÀÇ forward primer°¡ Æ÷ÇԵǾî ÀÖ¾î ÃÖ´ë 96°³ÀÇ illumina¢ç index Á¶ÇÕÀÌ °¡´ÉÇÏ´Ù. Library Á¦ÀÛ°ú Á¤Á¦¸¦ Æ÷ÇÔÇÑ NGS Library Àüü Á¦ÀÛÀº 6½Ã°£ À̳»¿¡ ¿Ï·áÇÒ ¼ö ÀÖ´Ù.

Alternate kits for other input amounts:


±×¸² 1. Schematic of technology in the SMARTer¢ç Stranded Total RNA-Seq Kit v2 - Pico Input Mammalian (³»¿ë ÀÚ¼¼È÷º¸±â)


±×¸² 2. SMARTer¢ç Stranded Total RNA-Seq Kit v2 - Pico Input MammalianÀ» ÀÌ¿ëÇÏ¿© Á¦ÀÛÇÑ NGS LibraryÀÇ ±¸Á¶.
The adapters added using 5' PCR Primer HT and 3' PCR Primer HT contain sequences allowing clustering on any Illumina flow cell (P7 shown in light blue, P5 shown in red), Illumina TruSeq¢ç HT indexes (Index 1 [i5] sequence shown in yellow, and Index 2 [i7] sequence shown in orange), as well as the regions recognized by sequencing primers Read Primer 2 (Read 2, purple) and Read Primer 1 (Read 1, green). Read 1 generates sequences antisense to the original RNA, while Read 2 yields sequences sense to the original RNA (orientation of original RNA denoted by 5' and 3' in dark blue). The first three nucleotides of the second sequencing read (Read 2) are derived from the Pico v2 SMART Adapter (shown as Xs). These three nucleotides must be trimmed prior to mapping if performing paired-end sequencing.


±×¸² 3. Cluster passing filter rate (%PF) Çâ»ó
Libraries generated with the Pico v1 or Pico v2 kits were pooled and run on NextSeq 500 or MiniSeq instruments, as indicated. For each graph, blue boxplots indicate the distribution of cluster densities for unfiltered (i.e., raw) reads, while the green boxplots indicate the distribution of cluster densities for reads that passed filtering. Quantities of reads passing filter (in millions) and %PF values for each sequencing run are included above each graph. The expected number of reads passing filter according to Illumina specifications was 130 million reads for runs on the NextSeq and 25 million reads for runs on the MiniSeq. Proportions of reads that aligned to PhiX sequences ranged from 0.5% to 1.15% for all sequencing runs. As indicated in the graphs, libraries generated with the Pico v2 kit achieved higher %PF values for both Illumina platforms relative to libraries generated with the Pico v1 kit, and yielded quantities of reads passing filter that greatly exceeded the Illumina specifications.


±×¸² 4. Version 1 Á¦Ç°°ú ºñ±³ÇÏ¿© ³ô¾ÆÁø Sequencing sensitivity¿Í Reproducibility
Sequencing libraries were generated from 1 ng and 10 ng inputs of total RNA extracted from human lung FFPE tissue using both the Pico v1 and Pico v2 kits, and sequenced on a NextSeq 500 instrument. Panel A. Sequencing metrics for libraries generated from 1 ng or 10 ng inputs using each kit. For both input amounts, the Pico v2 kit resulted in greater library yields, a lower proportion of reads mapping to rRNA and mtRNA, and a lower duplicate rate. For the 1 ng input, sequencing data from the Pico v2 library also identified thousands more transcripts than sequencing data from the Pico v1 library, indicating a higher sensitivity for Pico v2. Panel B. Comparison of transcript expression levels across input amounts. Higher reproducibility was observed between 1 ng and 10 ng inputs for data generated with the Pico v2 kit vs. data generated using the Pico v1 kit. FPKM values are shown on a Log10 scale. Transcripts represented in only one library can be seen along the X- and Y-axes of the scatter plots.
Àû¿ë
- Robust NGS library construction that retains strand information
- Use for RNA-seq on all Illumina platforms
- Captures coding and non-coding information from total mammalian RNA of any quality, including RNA obtained from FFPE samples
±¸¼ºÇ° (ÀÚ¼¼ÇÑ ±¸¼ºÇ°Àº CoA ¶Ç´Â ¸Å´º¾ó ÂüÁ¶)
- SMARTer Stranded Total RNA-Seq Kit v2 - Pico Input Mammalian Components
- Indexing Primer Set HT for Illumina v2

Keyword : NGS,RNA-Seq,Strand specific,Stranded-Seq,total RNA,ZapR,Illumina,Sequencing,½ÃÄö½Ì,ÀÏ·ç¹Ì³ª,SMART,SMARTer,smart-seq

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