[±â¼ú°³¿ä] ThruPLEX & PicoPLEX
DNA-Seq ±â¼ú°³¿ä
3 Steps, Single tube À̵¿ÀÌ ¾ø´Â ´Ü 3¹øÀÇ Á¶ÀÛÀ¸·Î Low-input DNA-seq Library Á¦ÀÛ
ThruPLEX¢ç technology
ThruPLEX¢ç ±â¼ú
(Step 1) ´ÜÆíÈµÈ double stranded DNA ¸»´ÜÀ» °íÈ¿À²·Î repairÇÑ´Ù.
(Step 2) DNA ÀýÆíÀÇ 5¡¯ ¸»´Ü¿¡ Stem-Loop Adapter°¡ ligation µÇ¸ç, 3¡¯ ¸»´Ü¿¡´Â nickÀ» ³²±ä´Ù. ÀÌÈÄ extension °úÁ¤À» ÅëÇØ Stem-Loop AdapterÀÇ ÀϺΰ¡ Á¦°ÅµÈ´Ù. Stem-Loop Adapter´Â ³ëÃâµÈ ´ÜÀÏ°¡´ÚÀÌ ¾ø°í 5¡¯ ¸»´ÜÀÌ blockµÇ¾î ÀÖ¾î adapter°£ÀÇ dimer Çü¼ºÀÌ ¾ïÁ¦µÉ »Ó¸¸ ¾Æ´Ï¶ó ¼¿ ºÐ¼®¿¡¼ÀÇ background ¸¦ °¨¼Ò½Ãų ¼ö ÀÖ´Ù.
(Step 3) Illumina
¢ç index°¡ Æ÷ÇÔµÈ primer·Î ÁõÆøÇÏ¿© Illumina
¢ç NGS ºÐ¼®¿ë library Á¦ÀÛÀ» ¿Ï·áÇÑ´Ù.
ThruPLEX¢ç ±â¼úÀÇ ÀåÁ¡
- ÇϳªÀÇ Æ©ºê¿¡¼ ´Ü 3 ¹øÀÇ Á¶ÀÛ, 15ºÐ hands-on-time
- Stem-loop adapter »ç¿ëÀ¸·Î background data °¨¼Ò
- ´Ù¾çÇÑ ºÐ¼® ½ÇÇè¿¡ ´ëÀÀ
- gDNA, cell-free DNA, FFPE DNA, cDNA, ChIP-DNA ¼¿ ºÐ¼®
- ´Ù¾çÇÑ target enrichment Ç÷§Æû ´ëÀÀ (Roche, Agilent ¿Ü)
Whole Genome Amplification (WGA)À» À§ÇÑ Gold Standard
PicoPLEX¢ç technology
PicoPLEX¢ç ±â¼ú
PicoPLEX
¢ç´Â quasi-random primer¸¦ ÀÌ¿ëÇÑ linear whole
genome pre-amplification °úÁ¤ (step 2)°ú exponential amplification
(step 3)À» ÀÌ¿ëÇÏ¿© ´ÜÀϼ¼Æ÷ (Single Cell) ¶Ç´Â ÇÇÄڱ׶÷ (pg) ¼öÁØÀÇ ±Ø¼Ò·®
DNA¸¦ ÁõÆøÇÏ°í ÀçÇö¼ºÀÌ ³ôÀº NGS library¸¦ ±¸ÃàÇÏ´Â ±â¼úÀÌ´Ù.
(Step 1) ¼¼Æ÷ ¿ëÇØ (cell lysis)
(Step 2) Pre-amplification
- µ¶ÀÚÀûÀÎ quasi-random primer°¡ gDNA¿¡ ¼±ÅÃÀûÀ¸·Î °áÇÕ
- PCR ¿¡·¯¸¦ ÃÖ´ëÇÑ °¨¼Ò½ÃÅ°´Â quasi-linear amplification Àû¿ë
(Step 3) PCR Amplification
PicoPLEX¢ç ±â¼úÀÇ ÀåÁ¡
- Single Cell ¶Ç´Â ±Ø¼Ò·® (6 pg ¼öÁØ) »ùÇÃÀ» ¸Å¿ì ³ôÀº °¨µµ¿Í ÀçÇö¼ºÀ¸·Î ÁõÆø ¹× ºÐ¼®
- À¯ÀüÀÚº¹Á¦º¯ÀÌ (CNV), ¿°»öü À̼ö¼º (Chromosomal aneuploidies), PGS/PGDÀÇ Gold standard
- 3 steps in a single tube