From: Charles Plessy <plessy@riken.jp>
Date: Thu, 19 Apr 2018 04:45:24 +0000 (+0900)
Subject: More details on Mg / Mn addition.
X-Git-Url: https://source.charles.plessy.org/?a=commitdiff_plain;h=9214d2093c09f79904e94c0911a15a937d6006da;p=source%2F.git

More details on Mg / Mn addition.
---

diff --git a/tags/template_switching.mdwn b/tags/template_switching.mdwn
index 7af520cb..f24e5178 100644
--- a/tags/template_switching.mdwn
+++ b/tags/template_switching.mdwn
@@ -12,10 +12,6 @@
    can extend a linker with the sequence of a small RNA via a template switching
    reaction.  (That is: a sRNA can play the same role as a TS oligonucleotide.)
 
- - [[Lee et al (2017)|biblio/28327113]] increased the efficiency of template
-   switching non-capped molecules by increasing dNTPs to 2 mM and
-   Mg<sup>2+</sup> to 9 mM.
-
 ### Effect of chemical composition of the TS oligonucleotide
 
 Originally, the TSOs were all-RNA.  Since this is expensive to synthesise,
@@ -33,4 +29,16 @@ as a replacement for RNA.
  - [[Arguel et al (2017)|biblio/27940562]] reported similar performance for
    RRR and RRL, using a 5′-focused method similar to nanoCAGE or STRT.
 
+### Effect of magnesium, manganese and dNTP concentrations
+
+ - [[Schmidt and Mueller, 1999|biblio/10518626]] showed that increasing
+   magnesium concentration (to 6 mM) or adding manganese at the end of the
+   reaction (1 or 2 mM) increased the frequency of dC addition (moderately
+   for Mg<sup>2+</sup> and strongly for Mn<sup>2+</sup>).  Enzyme: SSII; dNTP
+   concentration: 1 mM each.
+
+ - [[Lee et al (2017)|biblio/28327113]] increased the efficiency of template
+   switching non-capped molecules by increasing dNTPs to 2 mM and
+   Mg<sup>2+</sup> to 9 mM.
+
 [[!inline pages="tagged(template_switching)" limit=0]]