DoubleDigestion(雙酶切反應(yīng))時(shí)UniversalBuffer(通用緩沖液)的使用表

1、DoubleDigestion（雙酶切反應(yīng)）時(shí)UniversalBuffer（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)（DoubleDigestion）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UniversalBuffer表示系統(tǒng)，并對(duì)每種酶表示了在各UniversalBuffer中的相對(duì)活性。盡管如此，在進(jìn)行DoubleDigestion時(shí)，有時(shí)還會(huì)難以找到合適的UniversalBuffer。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DoubleDigestion可使用的最佳UniversalBuffer條件。在本表中，各Un

2、iversalBuffer之前表示的數(shù)字x是指各UniversalBuffer的反應(yīng)體系中的最終濃度oTaKaRa銷售產(chǎn)品中添附的UniversalBuffer全為10倍濃度的緩沖液。終濃度為0.5x時(shí)反應(yīng)體系中的緩沖液則稀釋至20倍，1x時(shí)稀釋至10倍,2x時(shí)稀釋至5倍進(jìn)行使用。注意1pgDNA中添加10U的限制酶，在50pl的反應(yīng)體系中，37C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，各DNA的立體結(jié)構(gòu)的差別，或當(dāng)限制酶識(shí)別位點(diǎn)鄰接時(shí)，有時(shí)會(huì)發(fā)生DoubleDigestion不能順利進(jìn)行的可能。Nde1xT1

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