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´Ù¾çÇÑ AAV serotypeÀÇ ºü¸£°í Á¤È®ÇÑ ¿ª°¡ ÃøÁ¤

Titration Kit (qPCR)

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Á¦Á¶»ç Á¦Ç°ÄÚµå Á¦Ç°¸í ¿ë·® °¡°Ý
(ºÎ°¡¼¼º°µµ)
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Takara
6233
AAVpro¢ç Titration Kit (for Real Time PCR)
°ü·ÃÇмú ±¸¸ÅÇϱ⠶óÀ̼±½º 
100 ȸ
682,000¿ø  6233_e.v2001.pdf





  • AAV2 ITR À¯·¡ÀÇ ´Ù¾çÇÑ AAV serotype Á¤·® °¡´É
  • ºü¸£°í °£ÆíÇÏ°Ô ¾à 2.5½Ã°£¸¸¿¡ AAV Á¤·® ¿Ï·á
  • ÀϹÝÀûÀÎ DNA blot ¶Ç´Â ELISA ¹æ¹ý ´ëºñ Á¤È®ÇÑ titration ÃøÁ¤ °¡´É
  • Packaging cell line¿¡¼­ AAV ÃßÃâ, Real Time PCR±îÁö ÇÊ¿äÇÑ ¸ðµç ±¸¼ºÇ° Æ÷ÇÔ
Á¦Ç° ¼³¸í
Adeno-Associated Virus (AAV)´Â ºÐ¿­, ºñºÐ¿­ ¼¼Æ÷¿¡ À¯ÀüÀÚ µµÀÔÀÌ °¡´ÉÇϸç, ƯÈ÷ ºñºÐ¿­ ¼¼Æ÷¿¡ ´ëÇØ Àå±â°£ À¯ÀüÀÚ ¹ßÇöÀÌ °¡´ÉÇÏ´Ù. ¶ÇÇÑ ¸é¿ª¿ø¼º (immune response)ÀÌ ³·¾Æ in-vivo ½ÇÇè¿¡ ÃÖÀûÈ­µÈ virusÀÌ´Ù.
AAVÀÇ ¿ª°¡ÃøÁ¤Àº ÀϰüµÈ ½ÇÇè °á°ú¿Í °¨¿° Á¶Á÷ ¹× °¨¿° ¹æ¹ý¿¡ ÃÖÀûÀÎ AAVÀÇ ¾çÀÇ °áÁ¤À» À§ÇØ ¸Å¿ì Áß¿äÇÏ´Ù.

1. AAVpro¢ç Titration Kit (for Real Time PCR) Ver.2 (Code 6233)

AAVpro¢ç Titration Kit (for Real Time PCR)Àº ´ÙÄ«¶ó°¡ µ¶ÀÚÀûÀ¸·Î °³¹ßÇÑ Real Time PCR¹æ½ÄÀÇ titration ¹æ¹ýÀ» »ç¿ëÇÏ¿©, ºü¸£°í È¿À²ÀûÀ¸·Î AAVÀÇ ¿ª°¡¸¦ ÃøÁ¤ÇÒ ¼ö ÀÖ´Ù. º» Á¦Ç°¿¡´Â AAV packaging cell lineÀ¸·ÎºÎÅÍ AAV¸¦ ÃßÃâÇÏ¿©, Real Time PCR Á¤·®À» À§ÇÑ ½Ã¾à±îÁö ¸ðµÎ Æ÷ÇԵǾî ÀÖ´Ù. AAVÀÇ vector ¾ç ¸»´Ü¿¡ Á¸ÀçÇÏ´Â ITR (Inverted Terminal Repeat)À» targetÀ¸·Î AAVÀÇ genomeÀ» °ËÃâÇϱ⠶§¹®¿¡, ±âÁ¸ÀÇ DNA blot ¶Ç´Â ELISA ¹ý¿¡ ºñÇØ ºü¸£°í Á¤È®ÇÑ °á°ú¸¦ ¾òÀ» ¼ö ÀÖÀ¸¸ç, ´Ù¾çÇÑ AAV serotype*1¿¡ Àû¿ëÇÒ ¼ö ÀÖ´Ù.
*1 AAV2À¯·¡ÀÇ ITR (ÀϹÝÀûÀ¸·Î »ç¿ë)À» °ËÃâÇϹǷÎ, ITR¿µ¿ªÀÌ AAV2·ÎºÎÅÍ À¯·¡ÇÑ ¼­¿­À̸é serotype¿¡ °ü°è¾øÀÌ Á¤·® °¡´ÉÇÏ´Ù.


±×¸² 1. AAVpro¢ç Titration Kit (for Real Time PCR) Ver.2 workflow


±×¸² 2. AAVpro¢ç Titration Kit (for Real Time PCR)ÀÇ Titer ÃøÁ¤ È¿À² ºñ±³.
AAV1, 2, 6¸¦ »ý»êÇÏ´Â packaging cell lineÀ¸·ÎºÎÅÍ AAVpro¢ç Extraction SolutionÀ» »ç¿ëÇÏ¿© AAV¸¦ ÃßÃâÇÏ¿´´Ù. ±× ÈÄ, AAVÀÇ ITR °ú CMV ºÎºÐÀ» °¢°¢ Ÿ°ÙÇÏ¿© Real Time PCRÀÇ Á¤·®°ªÀ» ºñ±³ÇÏ¿´´Ù (ITR: AAVpro¢ç Titration Kit (for Real Time PCR) Ver.2). º» µ¥ÀÌÅÍ´Â, AAVpro¢ç Titration Kit°¡ AAV2 ITR À¯·¡ÀÇ ´Ù¾çÇÑ serotype AAV Á¤·®¿¡ ÀÌ¿ë °¡´ÉÇÏ´Ù´Â °ÍÀ» ³ªÅ¸³½´Ù.

2. AAVpro¢ç Extraction Solution (Code 6235)
´ÙÄ«¶ó°¡ µ¶ÀÚÀûÀ¸·Î °³¹ßÇÑ AAV extraction½Ã¾àÀ¸·Î, ±âÁ¸ÀÇ freeze/thaw ¶Ç´Â sonication¹ý°ú °°ÀÌ ¹ø°Å·Î¿î °úÁ¤¾øÀÌ ºü¸£°í È¿À²ÀûÀ¸·Î AAV packaging cell lineÀ¸·ÎºÎÅÍ AAV¸¦ ÃßÃâÇÒ ¼ö ÀÖ´Ù (ƯÇã Ãâ¿ø Áß). º» Á¦Ç°Àº ´Ù¾çÇÑ serotypeÀÇ AAV¿¡ »ç¿ë °¡´ÉÇϸç, ¼÷ÁÖ (host) À¯·¡ ´Ü¹éÁú°ú ÇÙ»ê ¿À¿°ÀÌ ³·Àº AAV¸¦ ÃßÃâ °¡´ÉÇÏ´Ù.

[°ü·Ã Á¦Ç°]

<AAVpro¢ç Purification Kit>
ÀϹÝÀûÀÎ AAV Á¤Á¦ ¹æ¹ýÀÎCsCl2/iodixanol gradient ultracentrifugation ¹ýÀº °íµµÀÇ ¼÷·ÃµÈ ±â¼úÀ» ÇÊ¿ä·Î Çϸç, ¹ÙÀÌ·¯½º ȸ¼öÀ²ÀÌ ³·°í, ÃßÃâµÈ AAVÀÇ ³·Àº °¨¿°¼º°ú ½Ã°£ÀÌ ¿À·¡ ¼Ò¿äµÇ´Â ´ÜÁ¡ÀÌ ÀÖ´Ù. 
AAVpro¢ç Purification Kit¸¦ »ç¿ëÇÏ¸é ¾à 4½Ã°£¸¸¿¡ Æí¸®Çϰí, ºü¸£°Ô AAV¸¦ Á¤Á¦ ÇÒ ¼ö ÀÖ´Ù.
³»¿ë
AAVpro¢ç Titration Kit (for Real Time PCR) Ver.2 (Code 6233, 100 È¸)
1.

AAV ̧̉

 

AAV Extraction Solution A

1.5 §¢¡¿ 2

 

AAV Extraction Solution B

300 §¡

2.

AAV genome °£ÀÌÃßÃâ (100 »ùÇúÐ)

 

 

DNase I

100 §¡

 

10¡¿DNase I Buffer

200 §¡

 

Lysis Buffer

1 §¢¡¿ 2

3.

Real Time PCR (100 rxns: 25 ¥ìl ¹ÝÀÀ½Ã)

 

 

TB Green¢â Premix Ex Taq II

625 §¡¡¿ 2

 

AAV Forward Titer Primer

20 §¡

 

AAV Reverse Titer Primer

20 §¡

 

dH2O

1 §¢¡¿ 3

 

ROX Reference Dye

50 §¡

 

ROX Reference Dye II

50 §¡

 

Positive Control

50 §¡

 

EASY Dilution (for Real Time PCR)

1 §¢¡¿ 5


AAVpro¢ç Extraction Solution (Code 6235, 1 set)
1. AAV Extraction Solution A

25 §¢

2.

AAV Extraction Solution B

2.5 §¢

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AAVpro¢ç Titration Kit (for Real Time PCR) Ver.2 (Code 6233)

TB Green¢â Premix Ex Taq II: 4¡É, Â÷±¤ (Àå±â º¸Á¸½Ã -20¡É, À¶ÇØ ÈÄ¿¡´Â 4¡É º¸Á¸)
±× ¿Ü: -20 ¡É (AAV Extraction Solution A, B, Lysis Buffer´Â À¶ÇØ ÈÄ´Â ½Ç¿Â ÀúÀå)

AAVpro¢ç Extraction Solution (Code 6235) : ½Ç¿Â

Keyword : Adeno Associated Virus,in vivo,aav virus

Adeno-Associated Virus (AAV)
>
Adeno-associated virus Á¦ÀÛ¿¡ ÃÖÀûÀÎ
Packaging (293T Cell Line)
>
Helper Free ½Ã½ºÅÛ¿¡ ÀÇÇÑ ´Ù¾çÇÑ serotypeÀÇ ..
AAV1, 2, 5, 6 Vector System
>
Cre-Recombinase À¯ÀüÀÚ¸¦ žÀçÇÑ AAV1, AAV2, ..
CRE Recombinase System
>
¥â-galactosidase À¯ÀüÀÚ¸¦ žÀçÇÑ AAV1, AAV2,..
LacZ System
>
Çü±¤´Ü¹éÁúÀÌ Æ÷ÇÔµÈ AAV vector
pAAV-ZsGreen1 Vector
>
Adeno-associated virus delivery-CRISRP/Cas9 ..
CRISPR/Cas9 Vector System
>
SaCas9¸¦ ÀÌ¿ëÇÑ AAV mediated CRISPR Genome E..
CRISPR/SaCas9 Vector System (AAV2)
>
AAV2 Delivery Tet À¯µµ¹ßÇö ½Ã½ºÅÛ
Tet-Inducible System (AAV2)
>
´Ù¾çÇÑ serotypeÀÇ AAV Á¤Á¦¿¡ ÃÖÀû
Purification Kit Maxi/Midi (All Serotypes)
>
AAV Serotype 2 Á¤Á¦ kit
Purification Kit (AAV2 Àü¿ë)
>
AAV¸¦ È¿À²ÀûÀ¸·Î ³óÃà
AAVpro¢ç Concentrator
>
´Ù¾çÇÑ AAV serotypeÀÇ ºü¸£°í Á¤È®ÇÑ ¿ª°¡ Ãø..
Titration Kit (qPCR)
>
°£ÆíÇÑ AAV ÃßÃâ½Ã¾à
Extraction Solution