- ³ôÀº Á¤È®µµ: 99% ÀÌ»óÀÇ Á¤È®µµ·Î °¢ Àü»çüÀÇ origin °¡´Ú ½Äº° °¡´É
- ºü¸£°í °£¼ÒÈµÈ ÇÁ·ÎÅäÄÝ: ´Ü 5½Ã°£ ¸¸¿¡ library Á¦ÀÛ
- ³ÐÀº input ¹üÀ§: human, rat, mouseÀÇ total RNA 100 ng-1 ¥ìg
- ´Ù¾çÇÑ »ùÇà Àû¿ë¼º: RIN (RNA Integrity Number) °ªÀÌ 1-3 Á¤µµÀÇ total RNA¿¡¼µµ ³ôÀº ÀçÇö¼ºÀÇ °íÇ°Áú µ¥ÀÌÅ͸¦ »ý»ê
- High throughput Illumina sequencing: 384°³ÀÇ unique index¸¦ »ç¿ëÇÏ¿© ÃÖ´ë 384°³ÀÇ »ùÇ÷ΠIllumina-ready library Á¦ÀÛ °¡´É
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º» Á¦Ç°Àº 100 ng-1 §¶ÀÇ mammalian total RNA·ÎºÎÅÍ Illumina sequencing library¸¦ ÁغñÇϱâ À§ÇÑ È¿À²ÀûÀÎ ¼Ö·ç¼ÇÀ» Á¦°øÇÑ´Ù. ¶ÇÇÑ RiboGone¢â°ú SMART¢ç ±â¼úÀ» °áÇÕÇÏ¿© È¿À²ÀûÀ¸·Î rRNA¸¦ Á¦°ÅÇÏ°í strand ¹æÇ⼺ Á¤º¸°¡ À¯ÁöµÇ´Â libraryÀÇ ¿øÈ°ÇÑ »ý»êÀÌ °¡´ÉÇÏ´Ù.
rRNA´Â total RNAÀÇ ~90% ÀÌ»óÀ» Â÷ÁöÇϱ⠶§¹®¿¡ library »ý¼º Àü¿¡ rRNA°¡ È¿°úÀûÀ¸·Î Á¦°Å°¡ µÇ¾î¾ß sequencing ºñ¿ëÀ» ³·Ãß°í mapping statistics¸¦ Çâ»ó½Ãų ¼ö ÀÖ´Ù.
SMART¢ç ±â¼úÀ» ÀÌ¿ëÇÑ cDNA ÇÕ¼º °úÁ¤°ú PCRÀ» ÅëÇØ Unique Dual Index Kits (Code 634752-634756, º°µµ±¸¸Å)¿¡ ÀÖ´Â index ¹× adaptor¸¦ Ãß°¡ÇÏ°í, strand ¹æÇ⼺À» ºÎ¿©ÇÏ¿© °£´ÜÇÏ°Ô library prep ÇÁ·Î¼¼½º¸¦ ¿Ï·áÇÒ ¼ö ÀÖ´Ù. ¶ÇÇÑ rRNA Á¦°ÅºÎÅÍ Illumina¿ë index°¡ Ãß°¡µÈ RNA-Seq library Áغñ±îÁö ¾à 5½Ã°£ ³» ¿Ï¼ºµÈ´Ù.
*±âŸ input amount¿¡ µû¸¥ Ãßõ Á¦Ç° :
- Total RNA 250 pg-10 ng : SMARTer¢ç Stranded Total RNA-Seq Kit v3 - Pico Input Mammalian (Code 634485 ¿Ü)
- Total RNA 10-100 ng : SMART-Seq¢ç Total RNA Mid Input (Code 635049 ¿Ü)
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- Illumina Ç÷§ÆûÀ» ÀÌ¿ëÇÑ mammalian sampleÀÇ RNA-Seq
- Strand ¹æÇ⼺ Á¤º¸¸¦ À¯ÁöÇÏ´Â NGS library Áغñ
- coding ¹× noncoding ¼¿ Á¤º¸ ºÐ¼®
ÀÚ¼¼ÇÑ ±¸¼ºÀº CoA¸¦ Âü°íÇϼ¼¿ä.
RNA-Seq ·¯´×¼¾Å͸¦ ÅëÇØ º» Á¦Ç°¿¡ ´ëÇÑ ´õ ¸¹Àº µ¥ÀÌÅÍ¿Í FAQ¸¦ Âü°íÇϼ¼¿ä.