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AAV »ý»ê¿ë enhancer Ãß°¡ (GMP Ãâ½Ã)

VirusGEN¢ç AAV Transfection Kit

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Á¦Á¶»ç Á¦Ç°ÄÚµå Á¦Ç°¸í ¿ë·® °¡°Ý
(ºÎ°¡¼¼º°µµ)
ºñ°í »ç¿ëÀڸŴº¾ó
Mirus
MIR 6750
VirusGEN¢ç AAV Transfection Kit
°ü·ÃÇмú ±¸¸ÅÇϱâ
1 Kit (1 L cell culture)
(Package)
°¡°Ý¹®ÀÇ   virusgen-aav-transfection-kit-protocol.pdf
Mirus
MIR 6815-GMP
VirusGEN¢ç GMP AAV Transfection Kit
°ü·ÃÇмú ±¸¸ÅÇϱâ
1 Kit (50 L cell culture)
(Package)
°¡°Ý¹®ÀÇ   virusgen-gmp-aav-transfection-kit-protocol.pdf

VirusGEN® GMP kitÀÇ °ßÀû ¹× ±¸¸Å´Â ¹Ýµå½Ã ´ÙÄ«¶óÄÚ¸®¾Æ °í°´Áö¿ø (T. 02-2081-2510)À¸·Î ¹®ÀÇ ¹Ù¶ø´Ï´Ù.
AAV »ý»êÈ¿À²ÀÌ ´õ¿í ³ô¾ÆÁø ¾÷±×·¹À̵å Á¦Ç° VirusGEN¢ç AAV Transfection Kit with RevIT¢â AAV Enhancer (Code MIR 8007)À» È®ÀÎÇϼ¼¿ä!
  • ³ôÀº °¨¿°È°¼ºÀ» °¡Áö´Â °íÇ°ÁúÀÇ Adeno associated virus (AAV) »ý»ê¿¡ ÃÖÀû
  • HEK293 ¼¼Æ÷ ¹è¾ç ½Ã½ºÅÛ¿¡ ÃÖÀû (adherent ¹× suspension ¹è¾ç ¸ðµÎ Àû¿ë)
  • ¿¬±¸¿ë ½Ã¾àºÎÅÍ ÀüÀÓ»ó ´Ü°è, ÀÓ»ó¿ë GMP grade±îÁö ¶óÀξ÷
Á¦Ç°¼³¸í
TransIT-VirusGEN® Transfection Reagent´Â expression vector¿Í packaging plasmid¸¦ HEK293 ŸÀÔÀÇ ÆÐŰ¡ ¼¼Æ÷·Î È¿À²ÀûÀ¸·Î µµÀÔÇÏ¿©, AAV ¶Ç´Â LentivirusÀÇ »ý»êÀ²À» ³ô¿©ÁÖ´Â transfection ½Ã¾àÀÌ´Ù.
±× Áß¿¡¼­µµ º» VirusGEN® AAV Transfection Kit Á¦Ç°Àº TransIT-VirusGEN® Transfection Reagent¸¦ ±â¹ÝÀ¸·Î plasmid µµÀÔ È¿À²À» ³ôÀ̱â À§ÇÑ VirusGEN® AAV Complex Formation Solution and Enhancer°¡ Ãß°¡µÇ¾î AAVÀÇ È¿À²ÀûÀÎ »ý»êÀ» À§ÇØ ÃÖÀûÈ­µÈ Á¦Ç°ÀÌ´Ù.
VirusGEN® SELECT AAV Transfection Kit´Â ÀüÀÓ»ó ¿¬±¸ ¹× °øÁ¤ °³¹ß¿¡¼­ ÇÊ¿äÇÑ ´ë±Ô¸ð ¹ÙÀÌ·¯½º »ý»êÀ» À§ÇÑ ÃÖÀûÀÇ ½Ã¾àÀ¸·Î °£ÆíÇÏ°í ºñ¿ë È¿À²ÀûÀÎ ½ÇÇè ÇÁ·Î¼¼½º¸¦ Á¦°øÇÑ´Ù. ¶ÇÇÑ ÀüÀÓ»ó¿ë ¹ÙÀÌ·¯½º º¤ÅÍ Á¦ÀÛÀ» À§ÇÑ transfection ½Ã¾àÀ¸·Î »ç¿ëÇÒ ¼ö ÀÖµµ·Ï Ç°Áú º¸Áõ ½ÃÇèÀ» ¿Ï·áÇÏ¿´´Ù.
([Example Certificate of Origin (PDF) | Example Certificate of Analysis (PDF) | Example BSE TSE Statement (PDF)])
µû¶ó¼­, ÀÓ»ó ½ÃÇè ¶Ç´Â »ó¾÷ÀûÀÎ ¹ÙÀÌ·¯½º »ý»êÀ» À§ÇØ VirusGEN® GMP AAV Transfection Kit¸¦ »ç¿ëÇÒ ¼ö ÀÖ´Ù.


±×¸²1. Ÿ»ç´ëºñ ³ôÀº AAV functional titer
Suspension ÇüÅ·Π¹è¾çÇÑ Freestyle 293-F¼¼Æ÷ (FreeStyle F17 Medium¿¡¼­ ¹è¾ç)(A)¿Í Adherent ÇüÅ·Π¹è¾çÇÑ 293T/17¼¼Æ÷(B)¿¡ Trans IT-VirusGEN® Transfection Reagent (reagent : DNA (vol : wt) = 2 : 1), Ÿ»ç B4, Ÿ»ç B5, Ÿ»ç C3 (25kDa PEI)¸¦ »ç¿ëÇÏ¿© pAAV-hrGFP, pAAV-RC, pAAV-Helper (1 : 1 : 1 ratio, 1.5 §¶/§¢, Agilent Technologies)¸¦ transfection ÇÏ¿© ¹ÙÀÌ·¯½º¸¦ »ý»êÇÏ¿´´Ù. »ý»êµÈ ¹ÙÀÌ·¯½º´Â HT1080¼¼Æ÷¿¡ °¨¿°, 48½Ã°£ µÚ GFP¹ßÇöÀ» À¯¼¼Æ÷ÃøÁ¤±â·Î ÃøÁ¤ÇÏ¿´´Ù.
*Functional titer´Â GFP ¹ßÇö¼¼Æ÷°¡ 20% ¹Ì¸¸ÀÎ virus Èñ¼®¾×À¸·ÎºÎÅÍ ÃøÁ¤ÇÏ¿´°í, error bar´Â 3¹Ýº¹ ½ÇÇ豺ÀÇ Ç¥ÁØÆíÂ÷ÀÌ´Ù.


±×¸² 2. TransIT-VirusGEN® Reagent, VirusGEN® AAV Transfection Kit - Ÿ»ç Á¦Ç°°úÀÇ »ý»êÈ¿À² ºñ±³
TransIT-VirusGEN® Reagent¿Í VirusGEN® AAV Transfection Kit, Ÿ»ç A1, Ÿ»ç A3À» ÀÌ¿ëÇÏ¿© AAV2ÀÇ »ý»êÈ¿À²À» ºñ±³ÇÏ¿´´Ù. Expi293 Expression Media (Thermo Fisher Scientific)¿¡¼­ ¹è¾çÇÑ Expi293F cells (Thermo Fisher Scientific)¿¡ pAAV-hrGFP, pAAV-RC, pAAV-Helper plasmids (1:1:1 DNA ratio, 2 §¶/§¢ - 2 §· per flask, Agilent Technologies)¸¦ µµÀÔÇÏ°í 72½Ã°£ ÈÄ lysis ½ÃÄÑ AAV¸¦ ȸ¼öÇÏ¿´´Ù.

A. ȸ¼öÇÑ ¹ÙÀÌ·¯½º¸¦ HT10180¼¼Æ÷¿¡ °¨¿°½ÃÅ°°í 48½Ã°£ ÈÄ¿¡ GFPÀÇ ¹ßÇöÀ» È®ÀÎÇÏ¿© ¿ª°¡¸¦ ÃøÁ¤ÇÏ¿´´Ù. ¶Ç µµÀÔµÈ AAVÀÇ genome copy number´Â CMV promoter ¿µ¿ªÀ» ddPCR·Î È®ÀÎÇÏ¿´´Ù.
B. Á¦ÀÛµÈ ¹ÙÀÌ·¯½º capsid¼ö´Â AAV2 Titration ELISA Kit (Progen)À» ÀÌ¿ëÇÏ¿© ÃøÁ¤Çß´Ù. ¶Ç GC/capsidÀÇ ºñÀ²À» °è»êÇÏ¿´´Ù
±× °á°ú, TransIT-VirusGEN® Reagent, VirusGEN® AAV Transfection Kit¸¦ ÀÌ¿ëÇßÀ» ¶§ ´õ ¸¹Àº ¿ª°¡ÀÇ AAV¸¦ »ý»êÇÒ ¼ö ÀÖ¾ú°í, ƯÈ÷ VirusGEN® AAV Transfection Kit¸¦ ÀÌ¿ëÇÏ¿´À» ¶§ °¡Àå ¸¹Àº ¾çÀÇ AAV¸¦ »ý»êÇÒ ¼ö ÀÖ¾ú´Ù.
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TransIT-VirusGEN® Transfection Reagent: -10~-30¡É º¸Á¸
VirusGEN® AAV Complex Formation Solution and Enhancer : 2~10¡É
(SELECT, GMP ¸ðµÎ µ¿ÀÏ)
°ü·ÃÁ¦Ç°
- ´Ù¾çÇÑ SerotypeÀÇ vector system: AAVpro® Helper Free System (AAV1, 2, 5, 6)
- CRISPR/Cas9 Vector System: AAVpro® CRISPR/Cas9 Helper Free System (AAV2)
- AAV extraction: AAVpro® Extraction Solution
- Cell¿¡¼­ Á¤Á¦: AAVpro® Purification Kit (All Serotypes)
- ¹è¾ç¹èÁö¿Í cell¿¡¼­ ¸ðµÎ Á¤Á¦: AAVpro® Cell & Sup. Purification Kit Maxi (All Serotypes)
- AAV titration: AAVpro® Titration Kit (for Real Time PCR)
- Packaging (293T Cell Line): AAVpro® 293T Cell Line

Keyword :

È¿À²Áõ´ë¸¦ À§ÇÑ Æ¯¼öµµÀԽþà
>
°í¿ª°¡ÀÇ AAV, Lentivirus »ý»êÀ» À§ÇÑ (GMP Ãâ..
TransIT-VirusGEN¢ç Transfection Reagent
>
Recombinant Lentivirus »ý»ê¿¡ ÃÖÀû
TransIT¢ç-Lenti Transfection Reagent
>
AAV »ý»ê·®À» ³ôÀÌ´Â transfection enhancer
RevIT¢â AAV Enhancer
>
AAV »ý»ê¿ë enhancer Ãß°¡ (GMP Ãâ½Ã)
VirusGEN¢ç AAV Transfection Kit
>
Lentivirus »ý»ê¿ë enhancer Ãß°¡ (GMP Ãâ½Ã)
VirusGEN¢ç LV Transfection Kit