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Home > ÀüÁ¦Ç°º¸±â > Gene transfer (virus) > Retrovirus > Integration Site Analysis Kit

Integration Site Analysis Kit

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(ºÎ°¡¼¼º°µµ)
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Clontech
631467
Retro-X Integration Site Analysis Kit
°ü·ÃÇмú ±¸¸ÅÇϱâ
each
(Package)
1,458,400¿ø 
1,823,000¿ø
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11.01 ~ 12.27
Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â
x32401, x32402, x33008

  • GenomeWalker¸¦ ±â¹ÝÀ¸·Î provirusÀÇ °Ô³ð »ðÀÔ ºÎÀ§¸¦ ½Äº°
  • MMLV ¹× MSCVŸÀÔÀÇ retroviral vector¿¡ Àû¿ë °¡´É
  • ºÐ¼®¿¡ ÇÊ¿äÇÑ ¸ðµç ½Ã¾à Æ÷ÇÔ
Á¦Ç°¼³¸í
Retro-X Integration Site Analysis Kit´Â ·¹Æ®·Î ¹ÙÀÌ·¯½º º¤ÅÍ°¡ host genome DNA¿¡ provirus·Î »ðÀÔµÈ ºÎºÐÀ» GenomeWalker ±â¼úÀ» ±âºÐÀ¸·Î ºÐ¼®Çϱâ À§ÇÑ Á¦Ç°ÀÌ´Ù. ·¹Æ®·Î¹ÙÀÌ·¯½º´Â RNA genome virus·Î DNA·Î ¿ªÀü»ç µÇ¾î host genome¿¡ »ðÀԵȴÙ. ¹ÙÀÌ·¯½º DNA´Â host genome¿¡ ¹«ÀÛÀ§·Î »ðÀԵDZ⠶§¹®¿¡ ¸ñÀûÀ¯ÀüÀÚ ¹× endogenous À¯ÀüÀÚ ¹ßÇö¿¡ ¿µÇâÀ» ÁÙ ¼ö ÀÖ´Ù. µû¶ó¼­ provirusÀÇ »ðÀÔºÎÀ§ÀÇ ½Äº°Àº host cellÀÇ Ç¥ÇöÇü ¿¬±¸¿¡ Áß¿äÇÏ´Ù. ÀÌ Å°Æ®¿¡´Â ½ÇÇè¿¡ ÇÊ¿äÇÑ ¸ðµç ½Ã¾àÀÌ Æ÷ÇԵǾî ÀÖ´Ù. (ÀÚ¼¼ÇÑ ±¸¼ºÇ° ¸®½ºÆ®´Â CoA¸¦ Âü°í)
GenomeWalker °³¿ä
1. ¹ÙÀÌ·¯½º¸¦ ÇüÁú µµÀÔÇÑ ¼¼Æ÷¿¡¼­ ÃßÃâÇÑ genomic DNA¸¦ 3Á¾·ùÀÇ blunt-end Á¦ÇÑÈ¿¼Ò ó¸®
2. ÆòÈ°¸»´ÜÀÌ µÈ genomic DNA library¿¡ GenomeWalker adaptor¸¦ ligation.
3. Library¿¡¼­ adaptor¿Í ·»Æ¼¹ÙÀÌ·¯½º ½ÃÄý½º°¡ Æ÷ÇÔµÈ ºÎºÐÀ» PCR ÁõÆø
4. Nested PCRÀ» ÅëÇØ ´ÜÀϹêµå ÁõÆø, ÁõÆø µÈ »ê¹°Àº sequencingÀ» ÅëÇؼ­ integrationºÎºÐ ºÐ¼®, º¹¼öÀÇ ´ÜÆíÀÌ ÁõÆøµÇ´Â °æ¿ì¿¡´Â
    °¢°¢ÀÇ band¸¦ Á¤Á¦ ÇÑ ÈÄ sequencingÀ» ÅëÇؼ­ ºÐ¼®


±×¸². Flow chart of the Lenti-X Integration Site Analysis protocol. The major steps include construction of three viral integration libraries, PCR amplification of genomic DNA from the libraries, and analysis of PCR products. The gel shows a typical result. Lane 1: HpaI library. Lane 2: DraI library. Lane 3: SspI library. Lane 4: No template control. Lane M: DNA size markers. AP: Adaptor primer. LSP: Lentivirus-specific primer.
Components & Storage Conditions
°¢ Á¦Ç° ±¸¼º¹°°ú º¸°üÁ¶°ÇÀº Certificate of Analysis ¸¦ ÂüÁ¶ÇϽʽÿÀ.

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