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빠르고 정확한 AAV 역가측정 (qPCR)

AAVpro® Titration Kit (for Real Time PCR)

제조사	제품코드	제품명	용량	가격 (부가세별도)	비고	사용자매뉴얼
Takara	6233	AAVpro® Titration Kit (for Real Time PCR)	100 회	730,000원

Product index

다양한 serotype의 AAV를 정량
빠르고 간편하게 약5시간만에 AAV 정량
일반적인 DNA blot 또는 ELISA 방법 대비 정확한 titration 측정
Packaging cell line에서 AAV 추출, Real Time PCR까지 필요한 모든 구성품 포함

제품 설명

Adeno-Associated Virus (AAV)는 분열 및 비분열 세포에 모두 적용이 가능하며, 특히 비분열 세포에서 장기간 유전자 발현이 가능하다. 또한 면역원성 (immune response)이 낮아 in-vivo 실험에 최적화된 바이러스이다.
AAV의 역가측정은 감염 조직 및 감염 방법에 최적인 AAV의 양을 결정하고 일관된 실험 결과를 도출하기 위해 매우 중요하다. AAVpro^® Titration Kit (for Real Time PCR)은 다카라가 독자적으로 개발한 Real Time PCR방식의 titration Kit로 빠르고 효율적으로 AAV의 역가를 측정할 수 있다.

이 제품에는 AAV packaging cell line에서 AAV를 추출하는 과정부터 Real Time PCR 정량을 위한 시약이 모두 포함되어 있다. AAV vector 양 말단에 존재하는 ITR (Inverted Terminal Repeat)을 target으로 AAV genome을 검출하기 때문에, 기존의 DNA blot 또는 ELISA 법에 비해 빠르고 정확한 결과를 얻을 수 있으며, 다양한 AAV serotype^*1에 적용할 수 있다.
^*1 AAV에서 일반적으로 사용하는 AAV2유래의 ITR 서열을 검출하므로, ITR영역이 AAV2로부터 유래한 서열이면 serotype에 관계없이 정량 가능하다.

그림 1. AAVpro^® Titration Kit (for Real Time PCR) workflow

그림 2. 다양한 serotype에서 AAVpro^® Titration Kit (for Real Time PCR) 적용성 확인
ITR 서열 (AAVpro^® Titration Kit (for Real Time PCR)에서 사용)과, CMV의 서열을 타겟하는 각각의 primer set을 사용해 AAV1, 2, 6의 세 종류의 바이러스에 역가를 측정하였다. 그 결과 세 종류의 serotype 모두 CMV를 타겟한 결과와 유사한 경향을 보여 AAVpro^® Titration Kit가 AAV2유래의 ITR 서열을 가진 다양한 serotype의 AAV 정량에 이용 가능하다는 것을 확인했다.

내용

AAVpro^® Titration Kit (for Real Time PCR) (Code 6233, 100 회)

1.	AAV 추출 시약
	AAV Extraction Solution A	2 x 1.5 ㎖
	AAV Extraction Solution B	300 ㎕
2.	AAV genome 간이 추출 (100 회)
	DNase I	100 ㎕
	10×DNase I Buffer	200 ㎕
	Lysis Buffer	2 x 1 ㎖
3.	Real Time PCR (25 ㎕ 반응 시, 100회)
	TB Green™ Premix Ex Taq II (Tli RNaseH Plus)	2 x 625 ㎕
	AAV Forward Titer Primer	20 ㎕
	AAV Reverse Titer Primer	20 ㎕
	dH₂O	3 x 1 ㎖
	ROX Reference Dye	50 ㎕
	ROX Reference Dye II	50 ㎕
	Positive Control	50 ㎕
	EASY Dilution (for Real Time PCR)	5 x 1 ㎖

보존

TB Green^® Premix Ex Taq II (Tli RNaseH Plus): 4℃, 차광 (장기 보존시 -20℃, 융해 후에는 4℃ 보존)
그 외: -20 ℃ (AAV Extraction Solution A, B, Lysis Buffer는 융해 후는 실온 보존)

Citation

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2020 ASGCT Annual Meeting Abstracts
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Keyword : Adeno Associated Virus,in vivo,aav virus

Adeno-Associated Virus (AAV)

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CRISPR/SaCas9 Vector System (AAV2)

AAV2 Delivery Tet 유도발현 시스템

Tet-Inducible System (AAV2)

Cell과 배양 배지에서 AAV 추출부터 정제까지

Cell & Sup. Purification Kit

다양한 serotype의 AAV 정제에 최적

Purification Kit Maxi/Midi (All Serotypes)

AAV Serotype 2 정제 kit

Purification Kit (AAV2 전용)

AAV를 효율적으로 농축

AAVpro® Concentrator

빠르고 정확한 AAV 역가측정 (qPCR)

AAVpro® Titration Kit (for Real Time PCR)

간편한 AAV 추출시약

Extraction Solution

기존의 AAV 추출 방법으로 정제까지 4시간 내 ..

AAV Freeze-thaw extraction solution

AAVpro® Titration Kit (for Real Time PCR)

제품 설명

내용

보존

관련제품

Citation