Team:UT-Tokyo/Restriction Enzyme digestion（Xba1/Pst1）—Once for all

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<h1>Restriction Enzyme digestion（Xba1/Pst1）—Once for all</h1>

<h2>Preparation</h2>
*plasmid
*10×　buffer (H and M in the freezer)
*enzymeI,enzymeII（in the freezer in the next room）
*MilliQ
*1.5ml tube

<h3>■the correspondence of buffers</h3>
EX・・・M　ES・・・H　XP・・・M　SP・・・H

<h2>Procedure</h2>
<h3>■attention</h3>
*use 1.5ml tube
*enzyme must always be on ice! / Don’t leave at room temperature and heat!
put back in the freezer immediately!
*thaw frozen buffer fully! / you should spin down and vortex
*change MilliQ into new one everyday
※add MilliQ first and enzyme last / mix before add enzyme


<h3>for extracting gel</h3>
begin with 0.5μl enzyme and add to it if you worry (0.5μl is enough)
*1. add　?μl　plasmid（30μl for 1000ng）<br />
　　　　　3μl　10×　buffer<br />
　　　　　0.5μl　enzymeI<br />
　　　　　0.5μl　enzymeII<br />
　　　　　MilliQ　up to 30μl<br />
*2. incubate at 37 degrees for over 1hour<br />
(for 3 or 4 hour makes it sure)

<h3>for cut check</h3>
*1. add　1μl　plasmid (regardless of the concentration)<br />
　　　　　2μl　10×　buffer<br />
　　　　　0.5μl　enzymeI<br />
　　　　　0.5μl　enzymeII<br />
　　　　　MilliQ　up to 20μl<br />
*2. incubate at 37 degrees for over 1hour<br />
(for 3 or 4 hour makes it sure)

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