What is the duplicate rate in the final SMART-Seq Stranded cDNA library?

Duplicate rate depends on the input cell/RNA complexity and the sequencing depth. Low quantity cell/RNA inputs have lower complexity resulting in higher duplicate rates in the final SMART-Seq Stranded libraries. High sequencing depth will generate a higher duplicate rate, especially for low complexity samples.SMART-Seq Stranded [...]

How does SMART-Seq Stranded cDNA sequencing metrics correlate between RNA and cell inputs?

SMART-Seq Stranded cDNA synthesis shows a good sequencing metrics correlation between cells and purified RNA inputs.Libraries were generated using 10 pg of Control Total RNA (human brain) (Panels A and C) or a no-RNA control (Panel B) using the ultra-low workflow. Panel D shows an example of libraries generated directly from either single cells or a no-template control using the ultra-low [...]

What is the expected gene body coverage for the SMART-Seq Stranded library?

There is a slight 5’-end bias in the SMART-Seq Stranded cDNA, as is common for randomly primed libraries.SMART-Seq Stranded [...]

Can I use indexing primers from vendors other than Takara Bio USA?

Indexing primers from vendors other than Takara Bio USA, are not compatible with SMART-Seq Stranded Kit, since within the workflow of the SMART-Seq Stranded Kit, PCR incorporation of indexing primers requires proprietary overlaps with the first-strand cDNA template.SMART-Seq Stranded [...]

Do I need to use a PhiX DNA spike-in during the sequencing SMART-Seq Stranded cDNA libraries?

Illumina recommends the systematic inclusion of ~1% PhiX DNA to help assess run performance and troubleshooting. If performing paired-end sequencing on two-channel Illumina sequencer consider adding 5-10% PhiX.SMART-Seq Stranded [...]