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