雙酶切反應(yīng)通用緩沖液使用表

1、Double Digestion（ 雙酶切反應(yīng)）時(shí)Universal Buffer（ 通用緩沖液）的使用表說(shuō)明使用二種酶同時(shí)進(jìn)行 DNA切斷反應(yīng)（Double Digestion） 時(shí)，為了節(jié)省反應(yīng)時(shí)間，通常希望在同一反應(yīng)體系內(nèi)進(jìn)行。 TaKaRa采用 Universal Buffer 表示系統(tǒng)，并對(duì)每種酶表示了在各 Universal Buffer中的相對(duì)活性。盡管如此，在進(jìn)行 Double Digestion 時(shí)，有時(shí) 還會(huì)難以找到合適的 Universal Buffer 。本表以在pUC系列載體的多克隆位點(diǎn)處的各限制酶為核心，顯示了在 Double Digestion 可使用的最佳Uni

2、versal Buffer條件。在 本表中，各Universal Buffer之前表示的數(shù)字習(xí)是指各Universal Buffer的反應(yīng)體系中的最終濃度。TaKaRa銷(xiāo)售產(chǎn)品中添附的 Universal Buffer全為10倍濃度的緩沖液。終濃度為0.5用寸反應(yīng)體系中的緩沖液則稀釋至20倍，1 X0寸稀釋至10倍，2X時(shí)稀釋至5倍進(jìn)行使用。注息 1卬g DNA中添加10 U的限制酶，在50 "勺反應(yīng)體系中，37 C下反應(yīng)1小時(shí)可以完全降解 DNA。為防止Star活性的產(chǎn)生，請(qǐng)將反應(yīng)體系中的甘油含量，盡量控制在10%以下。根據(jù)DNA的種類(lèi)，各DNA的立體結(jié)構(gòu)的差別，或當(dāng)限制酶識(shí)別位點(diǎn)

3、鄰接時(shí)，有時(shí)會(huì)發(fā)生 Double Digestion 不能順利進(jìn)行的可 能。EnzymeAcc IBamH iBglWC!aEcoR 1EcoR VHine IIH/nd HIKpn 1Neo 1NdelAcc 1IxM0.5 xKIxT1xM1xM0.5 xK1xM1xM1xMIxMfBSAIxTfia/nH 10.5XK1XKIxK1 xK1 xK1 XK0.5 XK1 xK0.5 XKixK+BSAIxKBgl II1XT1XK1XH1 xH1 XH1 XH2XK1 xK1 XTIxKiBSA1xHClalIxM1XK1XH1 xM1 XH1 xH1 XM1XM1 xM1XK+BSA1

4、xH£coR IIxM1xK1XH1 XH1 XH1xH1 xM1XM1xMIxKfBSA1 xHfcoR V0.5 xKIxK1XH1 xH1 XH1 xH2xT1 XK0.5 XK1XK+BSA1 xHHine II1xM0.5 xK2xK1 XM1 xM2xT1 xM1 xM1xM1 乂瞄 SA1xTHind HlIxM1xKIXK1 xM1 xM1 xK1 xM1 xM1 xMixKtBSAIxKKpn 1IxM05 xK1XT1 XM1 xM0.5 xK1 xM1XM1 XL0.5XK48SA1xTNeo 1IxM+BSA1xK+BSAixK+BSAIxK曲IxK-hBSA

5、IxIGBSAIxMBSAIxKBSA0.5xK+BSAIxK+BSA1X KtBSANdel1XT1XK1XH1 XH1 xH1 XH1 XT1 XK1 XT1XK+8SA1xHNotl0.5xKfBSA0.5XK-BSAixHiBSAIxHiBSAixHBSAIXHBSA05XK.BSA05XK.BSAOSXK+BSA0.5XK+8SA1 x H.BSAPstl1xM1xK1XH1 XH1 XH1 XH1 XM1XM1xMIxK+BSA1 xHPvu I0.5 xKIxK1 x K1 XK1 XK1 xK0.5 KK1 xK0.5 XKIxK+BSAIxKSac 11XM05 XK0.5

6、XK1XM1XM0.5 XK1 XM1XM1 XLOixKf&SA1xTSai1.5x11.5XT1xH1 XH1 XH1 XH1.5XK1.5XK15XT+BSA15XT+BSA1 xHSma 1ixhBSA0.5XKBSA1XT+BSA1XT4BSA1XT4BSAOSxK+BSA1xT+BSA1XKBSAixT+BSA1XT+BSAIxTBSASpe 1IxMIxKIxH1xM1xH1xH1xM1xM1xMIxK+BSA1xHSph 10.5 xK1 xK1xH1 XH1 xH1 xH2xT1 xK0.5 XK1XK.8SA1 xHXb3IxM03 xK2xT1 XM1 XM2xT

7、1 xM1 xM1 XM1M8SA1xTXhoIxM1xK1xH1xH1xH1xH1xM1 xM1xM1xK1xHEnzymeNot IPstlPvuSacSa/1Sma 1SpelSph 1XbaXhoAcc0.5xKtBSA1xM0.5 xK1 xM1.5xT1 xTtBSA1 xM0.5 xK1xM1xMBamH I0.5XK 成A1xK1 xK0.5 xK1.5XT0.5XT+BSA1 xKIxK0.5XK1 xKBgll1XH+ESAIxH1 xK0.5 xK1xH1 xT+BSA1 xHIxH2xT1xHCla1XHfBSA1xH1 xK1 xM1 乂H1 xT+BSA1 xM1x

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