What is the expected double-stranded (ds) cDNA yield?
What is the expected double-stranded (ds) cDNA yield?
SMARTer Ultra low kits
The cDNA yield in newer generations of SMARTer Ultra low kits is higher compared to that of the legacy kits. Please refer to the user manual of your specific kit for more details. In general, depending on the RNA source, integrity, input amount, and the final volume of the library, the expected yield of ds cDNA generated using SMARTer Ultra low kits is 2–17 ng. This is achieved using the optimized number of PCR cycles and ensuring cDNA amplification is in the exponential phase (i.e., avoiding overcycling). To ensure true representation of the original mRNA pool, it is critical to avoid overamplification of cDNA. See ""Cycling Guidelines Based on Amount of Starting Material"" in the user manual for your particular SMARTer Ultra low kit.
Electropherogram of cDNA libraries generated with different amounts of Mouse Brain Total RNA (including a no RNA template control; NTC). Depending on the initial RNA input, optimization of PCR cycle number may be required to ensure a yield of 2–17 ng of cDNA.
SMARTer stranded kits
The SMARTer Stranded RNA-Seq Kit generates RNA-seq libraries for Illumina sequencing at a final concentration >7.5 nM. The SMARTer Stranded Total RNA Sample Prep Kit - HI Mammalian generates RNA-seq libraries for Illumina sequencing at a final concentration of 2–10 ng/µl.
SMARTer Universal Low Input RNA Kit for Sequencing
The expected yield of ds cDNA generated using the SMARTer Universal Low Input RNA Kit for Sequencing is 2–10 ng.