RRR and RRL, using a 5′-focused method similar to nanoCAGE or STRT.
- 3′ phosphate or biotin blocking groups abolish template-switching
- ([[Turchinovich et al (2014)|biblio/24922482]] and others).
+ ([[Turchinovich et al (2014)|biblio/24922482]] and others). However,
+ [[Pinto & Lindblad (2010)|biblio/19837043]] report the use of a
+ 3′ C3 spacer (on all-DNA TSOs).
- 5′ iso-dC and iso-dG prevents reverse-transcriptase to reach the end
of the TSO, and therefore blocks concatenation
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.
+ concentration: 1 mM each. [[Pinto & Lindblad (2010)|biblio/19837043]] also
+ used manganese.
- [[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]]