Team:UT-Tokyo/Restriction Enzyme digestion —One by one

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(Difference between revisions)
(New page: __NOTOC__{{UT-Tokyo_CSS3}}{{UT-Tokyo_Head}} <h1>Restriction Enzyme digestion —One by one</h1> <h2>Procedure（20μl）</h2> ! use 5ml tube<br /> 1. add　1ng　plasmid<br /> 　　　...)
 
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<h2>Procedure（20μl）</h2>
<h2>Procedure（20μl）</h2>
! use 5ml tube<br />
! use 5ml tube<br />
-
1. add　1ng　plasmid<br />
+
*1. add　1ng　plasmid
　　　　　2μl　10xbuffer<br />
　　　　　2μl　10xbuffer<br />
　　　　　2.5μl　enzyme<br />
　　　　　2.5μl　enzyme<br />
　　　　　MilliQ up to 20μl<br />
　　　　　MilliQ up to 20μl<br />
-
2. incubate at 37 degrees all night<br />
+
*2. incubate at 37 degrees all night<br />
<br />
<br />
Plasmid・MilliQ<br />
Plasmid・MilliQ<br />
-
EcoRI XbaI SpeI PstI<br />
+
EcoRI XbaI SpeI PstI
-
Buffer H M(+0.01%BSA) M H<br />
+
Buffer H M(+0.01%BSA) M H
<br />
<br />
buffer and activity of the restriction enzyme<br />
buffer and activity of the restriction enzyme<br />

Latest revision as of 07:44, 23 September 2010

UT-Tokyo

Restriction Enzyme digestion —One by one

Procedure（20μl）

! use 5ml tube

  • 1. add　1ng　plasmid

　　　　　2μl　10xbuffer
　　　　　2.5μl　enzyme
　　　　　MilliQ up to 20μl

  • 2. incubate at 37 degrees all night


Plasmid・MilliQ
EcoRI XbaI SpeI PstI Buffer H M(+0.01%BSA) M H
buffer and activity of the restriction enzyme
restriction enzyme relative activity（%）
L M H K EcoRI (20) (100) 100 (120) SpeI (80) 100 80 100 PstI (<20) (60) 100 80 XbaI <20 80* 20 <20

  • ＋0.01% BSA →100%
 Xba I

※（） tends to show the star activity

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