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Old 08-21-2014, 10:34 PM   #4
ecPDCNRA
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Location: Japan

Join Date: Oct 2011
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Default Concatemer synthesis from template switching oligo

FYI
The article shows the mechanism of concatemer synthesis from template switching oligo. Please see Figure 1 A. The authors show iso-G or iso-C nucleotide inhibit the concatemer synthesis.
>Incorporation of non-natural nucleotides into template-switching oligonucleotides reduces background and improves cDNA synthesis from very small RNA samples
http://www.biomedcentral.com/1471-2164/11/413


Quote:
Originally Posted by sylvain View Post
Hi everybody

A this time, we have done six Titanium run of cDNA sequencing. We always use SMART (SMARTER kit from Clontech) cDNA preparation without normalization. In the last run, we obtain at least 50% of SMART adaptor concatemer sequence.

Of course, we have followed the Clontech protocol and verified the quality and the amount of our RNA

Is anybody has an hypothesis to explain how this concatemer occur and how to modify the SMART original protocol in order to avoid this problem.

Thanks a lot
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