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Home > ÀüÁ¦Ç°º¸±â > NGS °ü·Ã > mRNA-Seq (Single cell) > [New]High throughput, Automation mRNA-Seq Kit
SMART-Seq mRNA ±â¹Ý, High-throughput mRNA-seq ºÐ¼®

[New]High throughput, Automation mRNA-Seq Kit

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Á¦Á¶»ç Á¦Ç°ÄÚµå Á¦Ç°¸í ¿ë·® °¡°Ý
(ºÎ°¡¼¼º°µµ)
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Clontech
634795
SMART-Seq¢ç mRNA HT
°ü·ÃÇмú ±¸¸ÅÇϱâ
24ȸ
(Package)
1,515,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â x345372
Clontech
634796
SMART-Seq¢ç mRNA HT
°ü·ÃÇмú ±¸¸ÅÇϱâ
96ȸ
(Package)
5,021,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â
Clontech
634791
SMART-Seq¢ç mRNA HT
°ü·ÃÇмú ±¸¸ÅÇϱâ
384ȸ (634796 x 4)
18,073,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â
Clontech
634792
SMART-Seq¢ç mRNA HT LP
°ü·ÃÇмú ±¸¸ÅÇϱâ
24ȸ
(Package)
2,024,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â x345377
Clontech
634793
SMART-Seq¢ç mRNA HT LP
°ü·ÃÇмú ±¸¸ÅÇϱâ
96ȸ
(Package)
7,070,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â
Clontech
634794
SMART-Seq¢ç mRNA HT LP
°ü·ÃÇмú ±¸¸ÅÇϱâ
384ȸ (634793 x 4)
25,452,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â

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Á¦Ç°»ç¿ë À¯¿¬¼ºÀ» À§ÇØ index°¡ kit¿¡ Æ÷ÇԵǾî ÀÖÁö ¾ÊÀ¸¸ç, º°µµ·Î ±¸¸Å °¡´ÉÇÕ´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
  • Easily automate the streamlined workflow - ªÀº ½Ã°£¿¡ ´Ù·®ÀÇ »ùÇÃÀ» ºÐ¼®ÇÒ ¼ö ÀÖÀ¸¸ç, ÀÚµ¿È­¿¡ ÃÖÀûÈ­µÈ ¿öÅ©Ç÷οì
  • Unparalleled sensitivity - Single cell ¶Ç´Â 10 pg total RNA »ùÇà ºÐ¼® °¡´É (Input range: ¼¼Æ÷ 1°³ - 100°³, ¶Ç´Â total RNA 10 pg - 1 ng)
  • High-quality RNA-seq data - Àü»çüÀÇ full-length coverage, rRNA read ÃÖ¼ÒÈ­, GC-rich Àü»çüÀÇ ±¤¹üÀ§ÇÑ representation
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  • Increase your sequencing power - ¸¹Àº ¼öÀÇ »ùÇÃÀ» Çѹø¿¡ ºÐ¼® ½Ã, Unique Dual Index (UDI)¸¦ »ç¿ëÇÏ¿© index hoping ¾øÀÌ multiplexing ºÐ¼® °¡´É
  • High library yield - NovaSeq¢â°ú °°Àº high-throughput sequencer¿¡ ½±°Ô Àû¿ëÀÌ °¡´ÉÇÏ°í, ³ôÀº yieldÀÇ library¸¦ Á¦ÀÛÇÒ ¼ö ÀÖ¾î ¿©·¯ Â÷·Ê ºÐ¼® °¡´É
°íÇ°ÁúÀÇ premium RNA
SMART-Seq¢ç mRNA HT¿Í SMART-Seq¢ç mRNA HT LP´Â oligo(dT) primer¸¦ »ç¿ëÇÏ¿© 10 pg - 1 ng total RNA ¶Ç´Â 1 - 100 °³ÀÇ ¼¼Æ÷¿¡¼­ Á÷Á¢ °íÇ°ÁúÀÇ full-length cDNA¸¦ ÇÕ¼ºÇÏ´Â ÀÚµ¿È­ ģȭÀûÀÎ kitÀÌ´Ù.
SMART-Seq¢ç mRNA HT¿¡´Â cDNA synthesis kit°¡, SMART-Seq¢ç mRNA HT LP¿¡´Â cDNA synthesis kitºÎÅÍ library preparation kit±îÁö Æ÷ÇԵǾî ÀÖ´Ù. Unique Dual Index kits¸¦ ÇÔ²² »ç¿ëÇϸé ÃÖ´ë 384°³ÀÇ multiplex Illumina-ready sequencing libraries¸¦ Á¦ÀÛÇÒ ¼ö ÀÖ´Ù.
SMART-Seq¢ç mRNA HT´Â FACS sorting ÀÌÈÄÀÇ ½ÇÇè ´Ü°è¸¦ ÃÖÀûÈ­ÇÏ°í, hands-on timeÀ» °¨¼Ò½ÃÅ´À¸·Î½á, Æí¸®¼ºÀ» ³ô¿´´Ù. cDNA¸¦ ÇÕ¼ºÇϴµ¥ »ç¿ëµÇ´Â SMART¢ç ±â¼úÀº full-lengthÀÇ Àü»çü Á¤º¸¸¦ Á¦°øÇÏ¿© splice junction ¹× alternative splicing Á¤º¸¸¦ ¾òÀ» ¼ö ÀÖ¾î single cellÀÇ gene expression ¿¬±¸¿¡ ¸Å¿ì À¯¿ëÇÏ´Ù. Oligo(dT) primingÀº °íÇ°Áú total RNA (RIN>8) ¶Ç´Â intact cells¿¡¼­ mRNA¸¦ ƯÀÌÀûÀ¸·Î ÁõÆøÇϴµ¥ »ç¿ëµÈ´Ù. LNA ±â¼úÀº template switching È¿°ú¸¦ Çâ»ó½ÃÄÑ ´Ù¸¥ ¹æ¹ýº¸´Ù ´õ ¸¹Àº À¯ÀüÀÚ Á¤º¸¸¦ ¾òÀ» ¼ö ÀÖ´Ù. ´ç»çÀÇ Æ¯Çã ¹ÞÀº library preparation chemistry ±â¼úÀ» Àû¿ëÇÑ SMART-Seq¢ç mRNA HT LP¿¡ Æ÷ÇÔµÈ library preparation kit´Â enzymatic fragmentation¿Í stem-loop adaptor¸¦ ÇÔ²² »ç¿ëÇÏ¿©, 2½Ã°£ ³» single tube¿¡¼­ purification °úÁ¤ ¾øÀÌ cDNA·ÎºÎÅÍ library·Î Á¦ÀÛÇÒ ¼ö ÀÖ´Ù. µû¶ó¼­ »ùÇÃÀÇ ¼Õ½ÇÀ̳ª ¿À¿° µî ½ÇÇè °úÁ¤¿¡¼­ ¹ß»ýÇÒ ¼ö ÀÖ´Â ¿À·ù¸¦ ÃÖ¼ÒÈ­ÇÒ ¼ö ÀÖ´Ù.

¸¸¾à degradationµÈ »ùÇÃÀ» °¡Áö°í Àְųª random priming ¹æ¹ýÀ» ¼±È£ÇÑ´Ù¸é SMART-Seq¢ç Stranded Kit¸¦ ÃßõÇÑ´Ù (index primer set¿Í library preparation kit°¡ Æ÷ÇԵǾî ÀÖÀ½).

Hads-on timeÀÌ Âª°í, °£¼ÒÈ­µÈ ¿öÅ©Ç÷ο츦 Á¦°øÇÏ´Â SMART-Seq¢ç mRNA HT


±×¸² 1. SMART-Seq¢ç mRNA kit¿Í SMART-Seq¢ç mRNA HT kitÀÇ ½ÇÇè°úÁ¤
SMART-Seq¢ç mRNA HT (Code 634795 ¿Ü) (¿À¸¥ÂÊ)´Â ±âÁ¸ÀÇ SMART-Seq¢ç mRNA (Code 634768 ¿Ü) (¿ÞÂÊ)ÀÇ ÇÁ·ÎÅäÄÝÀ» º¯ÇüÇÔÀ¸·Î½á º¸´Ù °£ÆíÇÏ°í ºü¸£°Ô High-throughputÀ¸·Î NGS Library¸¦ Á¦ÀÛÇÒ ¼ö ÀÖ´Ù. FACS sortingÀ¸·Î single cell ºÐ¸®ÇÑ ´ÙÀ½, SMART-Seq¢ç mRNA´Â ÃÑ 6 ´Ü°èÀÇ hands-on-stepÀÌ ÇÊ¿äÇÑ µ¥ ºñÇØ, SMART-Seq¢ç mRNA HTÀÇ °æ¿ì´Â ´Ü 3 ´Ü°èÀÇ hands-on-stepÀ¸·Î cDNA ÇÕ¼ºÀ» ¿Ï·áÇÒ ¼ö ÀÖ´Ù.

SMART-Seq¢ç mRNA (SMART-Seq¢ç v4ÀÇ ¸®´º¾ó Á¦Ç°)¿Í µ¿µîÇÑ ¼º´ÉÀ» °®´Â SMART-Seq¢ç mRNA HT

Sequencing metrics comparing SMART-Seq¢ç mRNA and SMART-Seq¢ç mRNA HT kits

RNA source

10 pg Mouse Brain Total RNA

cDNA synthesis

SMART-Seq¢ç mRNA

SMART-Seq¢ç mRNA HT

Replicate

A

B

C

A

B

C

Yield (ng)

7.3

8.5

9.4

9.5

9.9

9.2

Number of transcripts

14,168

13,934

14,147

14,510

14,650

14,553

11,455

11,267

11,349

11,582

11,670

11,454

Average Pearson/Spearman

0.97/0.67

0.97/0.68

0.96/0.66

Proportion of reads mapped (%)

rRNA

0.7

0.6

0.5

1.1

1.1

1.1

Mitochondria

2.8

4.2

4.1

3.7

3.8

4.2

Genome

92.3

90.8

86.6

89.1

89.0

87.8

Exons

74.8

72.5

69.0

67.3

67.1

65.3

Introns

13.3

14.1

13.3

16.9

16.9

17.4

Intergenic regions

4.2

4.2

4.4

4.9

5.0

5.2

Ç¥ 1. SMART-Seq¢ç mRNA¿Í SMART-Seq¢ç mRNA HT kit·Î °ËÃâÇÑ Àü»çüÀÇ ³ôÀº À¯»ç¼º
°¢ Á¦Ç°À» ÀÌ¿ëÇÏ¿© °¢°¢ mouse Brain Total RNA 10 pgÀ¸·Î library¸¦ Á¦ÀÛÇÏ¿© Àü»çü¸¦ °ËÃâÇÏ¿´°í, µÎ Á¦Ç°Àº ¸Å¿ì À¯»çÇÑ °á°ú¸¦ º¸¿´´Ù. cDNA »ê¹°Àº Illumina Nextera XT DNA Library Preparation Kit¸¦ ÀÌ¿ëÇÏ¿© RNA-Seq library·Î ÀüȯÇÏ°í Illumina NextSeq¢ç ±â±â (2 X 75bp)·Î ½ÃÄö½ÌÇÏ¿´´Ù.


±×¸² 2. Nextera XTº¸´Ù ´õ ³ôÀº library yield¿Í ´õ ¸¹Àº À¯ÀüÀÚ¸¦ ½Äº°ÇÏ´Â SAMRT-Seq¢ç mRNA HT LP
ÆгÎA. SMART-Seq¢ç mRNA HT·Î 10 pgÀÇ Mouse Brain Total RNA¿¡¼­ cDNA¸¦ ÇÕ¼ºÇÏ¿´´Ù (3ȸ ¹Ýº¹). ÀÌÈÄ SMART-Seq¢ç Library Prep (SSlp) ¶Ç´Â Nextera XT (Illumina) library prep kit·Î Illumina ȣȯ library¸¦ Á¦ÀÛÇÏ¿´´Ù. ±×¸®°í NextSeq 500 ±â±â·Î ½ÃÄö½ÌÇÏ¿´´Ù. Read´Â 4M paired-end reads·Î normalizationµÇ¾ú°í, ±â¼ú³ëÆ®¿¡ ¼³¸íµÈ ´ë·Î ºÐ¼®ÇÏ¿´´Ù. ÆгΠB. SSlp·Î ¾ò¾îÁø library yield´Â Nextera XT¿¡ ÀÇÇÑ yieldº¸´Ù ³ô¾Ò´Ù. ÆгÎC. SSlp´Â Nextera XTº¸´Ù 10% ´õ ¸¹Àº À¯ÀüÀÚ¸¦ ½Äº°ÇÒ ¼ö ÀÖ¾ú´Ù.
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- Àü»çü ½ÃÄö½ÌÀ» À§ÇÑ single cell ¶Ç´Â total RNA·ÎºÎÅÍÀÇ cDNA ÇÕ¼º°ú library Á¦ÀÛ
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ÀÚ¼¼ÇÑ ±¸¼ºÀº CoA¸¦ Âü°íÇϼ¼¿ä.
Á¦Ç°À» »ç¿ëÇϱâ Àü¿¡ Certificate of Analysis (CoA)¸¦ ÅëÇØ º¸°ü ¹æ¹ý°ú Á¦Ç° ±¸¼º, ±â¼úÀûÀÎ ¼º´ÉÀ» È®ÀÎÇϼ¼¿ä. Á¦Ç° ±¸¼ºÀ» È®ÀÎÇϱâ À§ÇØ »ç¿ëÀڸŴº¾óÀÇ ¡®List of Components¡¯¸¦ Âü°íÇϼ¼¿ä.

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