- ºü¸£°í °£´ÜÇÑ ÇÁ·ÎÅäÄÝ - Strand Á¤º¸ (sense/antisense)¸¦ °¡Áø Illumina¢ç sequencing-ready library¸¦ 7½Ã°£¸¸¿¡ Á¦ÀÛ
- Single cell ¼öÁرîÁö Àû¿ë °¡´É - 1-1,000 intact mammalian cell ȤÀº 10 pg - 10 ngÀÇ low/high-quality mammalian total RNA Àû¿ë
- ³ôÀº ÀçÇö¼º°ú Á¤È®µµ·Î ºÐ¼® - Single cell ¶Ç´Â total RNA·ÎºÎÅÍ full-lengthÀÇ coding/non-coding transcript¸¦ ÀçÇö¼º°ú Á¤È®µµ ³ô°Ô ºÐ¼®
- Single cell RNA-seqÀÇ gold standard SMART-Seq¢ç mRNA LP (with UMIs) (Code 634762 ¿Ü) ¼öÁØÀÇ ³ôÀº ¹Î°¨µµ
Á¦Ç°¼³¸í
SMART-Seq¢ç Total RNA Single Cell (ZapR¢â Mammalian)Àº random primer¸¦ »ç¿ëÇÏ¿© È¿°úÀûÀ¸·Î whole transcriptomeÀ» ºÐ¼®ÇÒ ¼ö ÀÖ´Â Á¦Ç°ÀÌ´Ù. ¶ÇÇÑ single cell ·¹º§¿¡¼ nonpolyadenylated transcripts¸¦ ºÐ¼®ÇÒ ¼ö ÀÖ´Ù. Stranded sequencing library Á¦ÀÛÀ» À§ÇØ cDNA ÇÕ¼º ¹× amplification ½Ã¾à, ZapR ±â¹Ý ribosomal RNA (rRNA) Á¦°Å½Ã¾à ±×¸®°í Illumina¢ç¿ë library Á¦À۽þàÀÌ Æ÷ÇԵǾî ÀÖ´Ù.
ÀÌ Á¦Ç°Àº ´ç»çÀÇ µ¶ÀÚÀûÀÎ SMART (Switching Mechanism at 5¡¯ end of RNA Template) ±â¼ú¿¡ stranded RNA-seqÀ» À§ÇÑ °³¼±»çÇ×À» ´õÇØ ÇÁ·ÎÅäÄÝÀ» °£ÆíÈÇÏ°í sequencing È¿À²À» ³ô¿´´Ù. ¶ÇÇÑ RT ´Ü°è¿¡¼ random primer¸¦ »ç¿ëÇØ nonpolyadenylated transcript¸¦ Æ÷ÇÔÇÏ´Â full transcriptome ºÐ¼®ÀÌ °¡´ÉÇÏ´Ù.
Single cell ¼öÁرîÁö Àû¿ë °¡´ÉÇÑ »ç¿ëÀÚ Ä£ÈÀûÀÎ °£´ÜÇÑ ÇÁ·ÎÅäÄÝ°ú rRNA À¯·¡ cDNA Á¦°Å ½Ã¾à (ZapR¢â mammalian)Àº °í°¨µµ·Î ÀçÇö¼º ³ôÀ¸¸ç rRNA mappingÀÌ ³·Àº µ¥ÀÌÅ͸¦ ¾òÀ» ¼ö ÀÖ´Ù. µû¶ó¼ º°µµÀÇ rRNA Á¦°Å kit ¾øÀÌ strand Á¤º¸ (sense /antisense)¸¦ °¡Áø sequencing library¸¦ °íÈ¿À²·Î Á¦ÀÛÇÒ ¼ö ÀÖ´Ù.
oligo dT primer¸¦ »ç¿ëÇÏ°íÀÚ ÇÑ´Ù¸é SMART-Seq¢ç mRNA LP (with UMIs) (Code 634762 ¿Ü) Á¦Ç°À» ÃßõÇÑ´Ù.
![](/file/manual/images/634360_01.jpg)
±×¸² 1. SMART-Seq¢ç Total RNA Single Cell (ZapR¢â Mammalian) ÇÁ·Î¼¼½º.
Àû¿ë
- ±Ø¼Ò·® (1-1,000) mammalian cell ¹× total RNA (10 pg - 10 ng)¿¡¼ 7½Ã°£¸¸¿¡ Illumina¢ç Ç÷§Æû¿ë sequencing library Á¦ÀÛ
- Strand Á¤º¸¸¦ °¡Áø whole transcriptome sequencing
ÀÚ¼¼ÇÑ ±¸¼ºÀº CoA¸¦ Âü°íÇϼ¼¿ä.
RNA-Seq ·¯´×¼¾ÅÍ¿¡¼ ´õ ¸¹Àº µ¥ÀÌÅÍ¿Í FAQ¸¦ È®ÀÎÇÏ½Ç ¼ö ÀÖ½À´Ï´Ù.