Can anyone offer any insight why in SMART-seq2 the RT and TSO primers are designed in a way that cDNA is amplified by semi-suppressive PCR (suggesting that without it, there would be a lot of primer-dimer products amplified) whereas in SMART-seq3/FLASH-seq, the RT/TSO primers carry different anchor sequences for cDNA amplification which does not rely anymore on semi-suppressive nature. So is the primer-dimer formation suppressed by either different RT enzyme usage or reaction buffer conditions in SS3/FLASH-seq or somehow these new RT/TSO primers do not form primer-dimers as efficiently as those in SMART-seq2? Is there then any RT/TSO design principle that one should follow to avoid excess formation of primer-dimers during template-switching RT? Thanks a lot!
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Researchers using sequencing and genomics tools often have to make trade-offs. They can choose between speed or scale, short reads or long-range information, or targeted panels or a view of the whole transcriptome. New technologies that have been released this year are built to address those tough choices.
We asked six companies the same four questions to learn about their latest products. The new technologies bring a lot to the table, including rethinking sequencing...
