Table 1 Summary of the effects of reaction conditions on the yDcpS activity.

From: The yeast scavenger decapping enzyme DcpS and its application for in vitro RNA recapping

Supplemented Salt

% decapped

Buffer, pH

% decapped

0 mM NaCl

>95

NaOAc pH 5.5

33

5 mM NaCl

>95

NaOAc pH 6

79

50 mM NaCl

>95

Bis-Tris pH 6.5

82

100 mM NaCl

42

K phosphate pH 7

16

200 mM NaCl

6

Tris-HCl pH 7.5

35

5 mM KCl

>95

Tris-HCl pH 8

13

50 mM KCl

72

Tris-HCl pH 8.5

<5

100 mM KCl

20

VCE Buffer pH 8

<5

200 mM KCl

<5

  
  1. In vitro decapping of 5′-capped synthetic 3′-FAM-labeled RNA under different reaction conditions at 60 nM yDcpS. All reactions contained an additional 30 mM NaCl because of the contribution from the enzyme storage buffer. The left column shows the effect of salt on the reaction. The yDcpS reaction buffer, which is 10 mM Bis-Tris pH 6.5 and 1 mM EDTA, was supplemented with the indicated concentration of salt. The right column shows the effect of pH on the reaction. The concentration of each buffering agent was 10 mM and contained 1 mM EDTA. The VCE Buffer is from NEB. See Methods section for more details.
Back to article page