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Home > 전제품보기 > 제한효소 · Quickcut > 제한효소 > Double Digestion용 추천 Universal Buffer

Double Digestion용 추천 Universal Buffer

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Double digestion buffer 선택가이드

두 종의 제한효소를 동시에 사용하여 DNA를 절단하는 double digestion 실험은 시간을 절약하기 위해 종종 수행된다. 당사에서 판매하는 제한효소에는 universal buffer가 포함되어 있지만 (각 buffer에 따른 상대 활성은 ‘Universal buffer별 활성’ 페이지 참조), 이 중 일부 double digestion의 경우, 두 효소에 모두 적절한 buffer를 선택하는 것이 어려울 수 있다. 각기 다른 두 효소의 높은 활성은 유지하고 star activity를 방지하기 위해 적절한 buffer를 선택하는 것이 중요하다.
일반적인 double digestion에 적절한 buffer, 희석배수 및 필수 첨가시약을 아래의 표에서 확인할 수 있다.
L, M, H, T, K는 buffer 유형을 나타내고, 권장되는 최종 buffer 농도로 표시된다 (universal buffer는 10X 농도로 제공됨). BSA도 10X 농도로 공급되므로, 10배 희석하여 최종 농도 0.01%로 사용한다.
단, 아래의 표에서 확인되지 않는 조합의 경우, double digestion을 대신 각 제한효소의 조건에 따라 따로 처리할 것을 권장한다.

Enzyme

Acc I

BamH I

Bgl II

Cla I

EcoR I

EcoR V

Hinc II

Hind III

Kpn I

Nco I

Nde I

Supplied
Buffer

10X M

10X K

10X H

10X M

10X H

10X H

10X M

10X M

10X L

10X K
+ BSA

10X H

Acc I

-

0.5X K

1X T

1X M

1X M

0.5X K

1X M

1X M

1X M

1X M
+ BSA

1X T

BamH I

0.5 × K

-

1X K

1X K

1X K

1X K

0.5X K

1X K

0.5X K

1X K
+ BSA

1X K

Bgl II

1X T

1X K

-

1X H

1X H

1X H

2X K

1X K

1X T

1X K
+ BSA

1X H

Cla I

1X M

1X K

1X H

-

1X H

1X H

1X M

1X M

1X M

1X K
+ BSA

1X H

EcoR I

1X M

1X K

1X H

1X H

-

1X H

1X M

1X M

1X M

1X K
+ BSA

1X H

EcoR V

0.5X K

1X K

1X H

1X H

1X H

-

2X T

1X K

0.5X K

1X K
+ BSA

1X H

Hinc II

1X M

0.5X K

2X K

1X M

1X M

2X T

-

1X M

1X M

1X M
+ BSA

1X T

Hind III

1X M

1X K

1X K

1X M

1X M

1X K

1X M

-

1X M

1X K
+ BSA

1X K

Kpn I

1X M

0.5X K

1X T

1X M

1X M

0.5X K

1X M

1X M

-

0.5X K
+ BSA

1X T

Nco I

1X M
+ BSA

1X K
+ BSA

1X K
+ BSA

1X K
+ BSA

1X K
+ BSA

1X K
+ BSA

1X M
+ BSA

1X K
+ BSA

0.5X K
+ BSA

-

1X K
+ BSA

Nde I

1X T

1X K

1X H

1X H

1X H

1X H

1X T

1X K

1X T

1X K
+ BSA

-

Not I

0.5X K
+ BSA

0.5X K
+ BSA

1X H
+ BSA

1X H
+ BSA

1X H
+ BSA

1X H
+ BSA

0.5X K
+ BSA

0.5X K
+ BSA

0.5X K
+ BSA

0.5X K
+ BSA

1X H
+ BSA

Pst I

1X M

1X K

1X H

1X H

1X H

1X H

1X M

1X M

1X M

1X K
+ BSA

1X H

Pvu I

0.5X K

1X K

1X K

1X K

1X K

1X K

0.5X K

1X K

0.5X K

1X K
+ BSA

1X K

Sac I

1X M

0.5X K

0.5X K

1X M

1X M

0.5X K

1X M

1X M

1X L

0.5X K
+ BSA

1X T

Sal I

1.5X T

1.5X T

1X H

1X H

1X H

1X H

1.5X K

1.5X K

1.5X T
+ BSA

1.5X T
+ BSA

1X H

Sma I

1X T
+ BSA

0.5X T
+ BSA

1X T
+ BSA

1X T
+ BSA

1X T
+ BSA

0.5X K
+ BSA

1X T
+ BSA

1X T
+ BSA

1X T
+ BSA

1X T
+ BSA

1X T
+ BSA

Spe I

1X M

1X K

1X H

1X M

1X H

1X H

1X M

1X M

1X M

1X K
+ BSA

1X H

Sph I

0.5X K

1X K

1X H

1X H

1X H

1X H

2X T

1X K

0.5X K

1X K
+ BSA

1X H

Xba I

1X M

0.5X K

2X T

1X M

1X M

2X T

1X M

1X M

1X M

1X M
+ BSA

1X T

Xho I

1X M

1X K

1X H

1X H

1X H

1X H

1X M

1X M

1X M

1X K
+ BSA

1X H


Enzyme

Not I

Pst I

Pvu I

Sac I

Sal I

Sma I

Spe I

Sph I

Xba I

Xho I

Supplied
Buffer

10X H
+ BSA 
+ Triton

1X H

10×K
+ BSA

10X L

10X H

10X T
+ BSA

10X M

10X H

10X M
+ BSA

10 X H

Acc I

0.5X K
+ BSA

1X M

0.5X K

1X M

1.5X T

1X T
+ BSA

1X M

0.5X K

1X M

1X M

BamH I

0.5X K
+ BSA

1X K

1X K

0.5X K

1.5X T

0.5X T
+ BSA

1X K

1X K

0.5X K

1X K

Bgl II

1X H
+ BSA

1X H

1X K

0.5X K

1X H

1X T
+ BSA

1X H

1X H

2X T

1X H

Cla I

1X H
+BSA

1X H

1X K

1X M

1X H

1X T
+BSA

1X M

1X H

1X H

1X H

EcoR I

1X H
+ BSA

1X H

1X K

1X M

1X H

1X T
+ BSA

1X H

1X H

1X M

1X H

EcoR V

1X H
+ BSA

1X H

1X K

0.5X K

1X H

0.5X K
+ BSA

1X H

1X H

2X T

1X H

Hinc II

0.5X K
+ BSA

1X M

0.5X K

1X M

1.5X K

1X T
+ BSA

1X M

2X T

1X M

1X M

Hind III

0.5X K
+ BSA

1X M

1X K

1X M

1.5X K

1X T
+ BSA

1X M

1X K

1X M

1X M

Kpn I

0.5X K
+BSA

1X M

0.5X K

1X L

1.5X T
+BSA

1X T
+BSA

1X M

0.5X K

1X M

1X M

Nco I

0.5X K
+ BSA

1X K
+ BSA

1X K
+ BSA

0.5X K
+ BSA

1.5X T
+ BSA

1X T
+ BSA

1X K
+ BSA

1X K
+ BSA

1X M
+ BSA

1X K
+BSA

Nde I

1X H
+ BSA

1X H

1X K

1X T

1X T
+ BSA

1X H

1X H

1X T

1X H

1X H

Not I

-

1X H
+ BSA

2X K
+ BSA

0.5X K
+ BSA

1X H
+ BSA

0.5X T
+ BSA

1X H
+ BSA

1X H
+ BSA

0.5X K
+ BSA

1X H
+BSA

Pst I

1X H
+ BSA

-

1X K

1X M

1X H

0.5X T
+ BSA

1X H

1X H

1X M

1X H

Pvu I

2X K
+ BSA

1X K

-

0.5X K

1.5X K
+ BSA

1X K
+ BSA

1X K

1X K

0.5X K

1X K

Sac I

0.5X K
+ BSA

1X M

0.5X K

-

1.5X T
+ BSA

1X T
+ BSA

1X M

0.5X K

1X M

1X M

Sal I

1X H
+ BSA

1X H

1.5X K
+ BSA

1.5X T
+ BSA

-

1.5X T
+ BSA

1X H

1X H

1.5X T

1X H

Sma I

0.5X T
+ BSA

0.5X T
+ BSA

1X K
+ BSA

1X T
+ BSA

1.5X T
+ BSA

-

1X T
+ BSA

0.5X T
+ BSA

1X T
+ BSA

1X T
+BSA

Spe I

1X H
+ BSA

1X H

1X K

1X M

1X H

1X T
+ BSA

-

1X H

1X M

1X H

Sph I

1X H
+ BSA

1X H

1X K

0.5X K

1X H

0.5X T
+ BSA

1X H

-

2X T

1X H

Xba I

0.5X K
+ BSA

1X M

0.5X K

1X M

1.5X T

1X T
+ BSA

1X M

2X T

-

1X M

Xho I

1X H
+ BSA

1X H

1X K

1X M

1X H

1X T
+ BSA

1X H

1X H

1X M

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Notes:
각 효소 10 unit은 50 ㎕ 반응액에서 37℃, 1 시간 내에 1 ㎍의 DNA를 완전히 분해한다.
Star activity를 최소화하려면 glycerol 농도가 10% 미만이어야 한다.
고차 구조의 DNA는 두 제한효소의 인식서열이 서로 가까울 경우 DNA가 완전히 절단되지 않을 수 있다.