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Y2H Library Construction System

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Á¦Á¶»ç Á¦Ç°ÄÚµå Á¦Ç°¸í ¿ë·® °¡°Ý
(ºÎ°¡¼¼º°µµ)
ºñ°í »ç¿ëÀڸŴº¾ó
Clontech
630490
Make Your Own ¡°Mate & Plate¡± Library System
°ü·ÃÇмú ±¸¸ÅÇϱâ LMOÁ¦Ç° 
5 ȸ
(Package)
3,120,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â x33366, x32807, x33213

  • SMART ±â¼úÀ» »ç¿ëÇÏ¿© yeast¿¡¼­ Á÷Á¢ library Á¦ÀÛ
  • ¾î·Á¿î cloningÀ̳ª library ÁõÆø °úÁ¤ ºÒÇÊ¿ä
  • Yeast two-hybrid screeningÀ» ¼ö¹éȸ ÇÒ ¼ö ÀÖÀ» ¸¸Å­ ÃæºÐÇÑ ½Ã¾àÀ¸·Î ±¸¼ºµÊ
Mate & Plate ÀåÁ¡
ÀüÇüÀûÀÎ ¹æ¹ýÀ¸·Î yeast two-hybrid¿ë library¸¦ Á¦ÀÛÇÏ°í screening ÇÏ´Â °ÍÀº ³ëµ¿ Áý¾àÀûÀÎ °úÁ¤ÀÌ´Ù. ÇÏÁö¸¸, Mate & Plate Library´Â ¹Ì¸® library·Î ÇüÁú ÀüȯµÈ yeast strain (MAT¥á)À¸·Î »õ bait ´Ü¹éÁúÀ» ¹ßÇöÇÏ´Â reporter strain (MAT¥á)°ú ÇÔ²² ÇÏ·í¹ã ¹è¾çÇÏ¿© ÀûÀýÇÑ selective/minimal ¹èÁö¿¡ µµ¸»ÇÏ¸é µÇ¹Ç·Î, ¾ÆÁÖ ½±°Ô »óÈ£ÀÛ¿ëÀ» Ž»öÇÒ ¼ö ÀÖ´Ù. ClontechÀº library screening¿¡ Àû¿ë °¡´ÉÇÑ ´Ù¾çÇÑ ÇüÅÂÀÇ tissue-specific library, normalized library ¹× universal library¸¦ Á¦°øÇÏ°í ÀÖ´Ù.


Figure 1. Library generation using in vivo homologous recombination in yeast. Mate & Plate Libraries are created via recombination between your SMART-generated cDNA and the Matchmaker prey vector, pGADT7-Rec. Transformed yeast colonies are pooled, mixed, and aliquoted into multiplevials. Each single 1 ml vial can be used for a two-hybrid screen.
°£´ÜÇÏ°í ºü¸£°Ô ¸¸µé ¼ö ÀÖ´Â ÀڽŸ¸ÀÇ Library
ClontechÀÇ Mate & Plate Library¿¡¼­ ÀÚ½ÅÀÇ ¸ñÀû¿¡ ÀûÇÕÇÑ Library¸¦ ãÀ» ¼ö ¾ø´Ù¸é, Make Your Own Mate & Plate Library SystemÀ» »ç¿ëÇÏ´Â °Íµµ ÁÁ´Ù. º» systemÀ» »ç¿ëÇÏ¸é ¼ö ¹éȸÀÇ yeast two-hybrid screeningÀ» ÇÒ ¼ö ÀÖÀ» ¸¸Å­ ÃæºÐÇÑ ¾çÀÇ library¸¦ ÀÏÁÖÀÏ ¾È¿¡ Á¦ÀÛÇÒ ¼ö ÀÖ´Ù. Library Á¦ÀÛÀº yeast ÀÚüÀÇ homologous recombination Çö»óÀ» »ç¿ëÇÏ¿© Y187 library yeast strain¿¡¼­ Á÷Á¢ ÀÌ·ç¾îÁø´Ù (Figure 1). ÀϹÝÀûÀÎ library Á¦ÀÛ °úÁ¤¿¡ ÇÊ¿äÇÑ ¼ö°í (¿¹: library cloning, ÁõÆø, Á¤Á¦ µî)¸¦ ÇÇÇÒ ¼ö ÀÖ´Ù.
°æÁ¦ÀûÀÎ SMART cDNA ÇÕ¼º ±â¼ú
º» systemÀº ClontechÀÇ SMART cDNA ÇÕ¼º ±â¼úÀ» »ç¿ëÇÏ¿© total RNA 100 ng Á¤µµÀÇ ÀûÀº ¾çÀ¸·Î ¾î¶°ÇÑ Á¶Á÷ÀÇ cDNA libraryµµ Á¦ÀÛÇÒ ¼ö ÀÖ´Ù. SMART ±â¼úÀº Moloney murine leukemia virus ¿ªÀü»çÈ¿¼ÒÀÇ terminal transferase¿Í template switching activity¸¦ ÀÌ¿ëÇÏ¿© ù ¹ø°·Î ÇÕ¼ºµÇ´Â cDNA ¾ç ¸»´Ü¿¡ ¼­¿­À» ¾Ë°í ÀÖ´Â primer binding site°¡ »ý¼ºµÇµµ·Ï ÇÑ´Ù.µû¶ó¼­ SMART ±â¼ú·Î ÇÕ¼ºµÈ cDNA´Â ¹Ù·Î PCR ÁÖÇüÀ¸·Î »ç¿ëÀÌ °¡´ÉÇÏ´Ù. ¶ÇÇÑ, ÀÌ·¸°Ô ÁõÆøµÈ cDNA´Â homologous recombinationÀ» ÀÌ¿ëÇÏ¿© Matchmaker Gold prey plasmidÀÎ pGADT7-RecÀ¸·Î cloningÇÒ ¼ö ÀÖ´Ù. ÀÌ·¯ÇÑ Æ¯¼ºÀ¸·Î ÀÎÇÏ¿© ng ¼öÁØÀÇ RNA (microdissected tissues, lasercaptured cells, biopsy samples µî)·ÎºÎÅÍ cDNA¸¦ ÇÕ¼ºÇÏ¿© pGADT7-Rec vector¿Í ÇÔ²² yeast strain Y187·Î co-transformationÇÏ¿© Á÷Á¢ÀûÀ¸·Î library¸¦ Á¦ÀÛÇÒ ¼ö ÀÖ´Ù.
Components & Storage Conditions
°¢ Á¦Ç° ±¸¼º¹°°ú º¸°üÁ¶°ÇÀº Certificate of Analysis ¸¦ ÂüÁ¶ÇϽʽÿÀ.

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