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Home > ÀüÁ¦Ç°º¸±â > Epigenetics > ChIP-seq, Cut&Run-seq (NGS) > [DNA-Seq ±â¼ú°³¿ä] ThruPLEX & PicoPLEX

[DNA-Seq ±â¼ú°³¿ä] ThruPLEX & PicoPLEX

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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

SMARTer¢ç ThruPLEX¢ç technology




SMARTer¢ç 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.



SMARTer¢ç 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

SMARTer¢ç PicoPLEX¢ç technology


SMARTer¢ç PicoPLEX¢ç ±â¼ú

SMARTer¢ç 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


SMARTer¢ç 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