The SMARTer Stranded Total RNA-Seq Kit v3 - Pico Input Mammalian is used to generate strand-specific RNA-seq libraries for Illumina sequencing from 250 pg–10 ng inputs of purified total RNA or from 10–1,000 intact cells. This kit incorporates Takara Bio’s proprietary SMART (Switching Mechanism at the 5’ end of RNA Template) technology and includes refinements to the SMARTer method for stranded RNA-seq that simplify the library preparation workflow and improve sequencing performance. This method was developed to work with either high- or low-quality total RNA, does not require additional rRNA removal methods or kits, and produces sequencing libraries that retain strand-of-origin information. The integrated removal of cDNAs derived from rRNA—typically present in high abundance following cDNA synthesis from total RNA inputs—makes the workflow extremely sensitive, yielding data that is highly reproducible with low mapping to rRNA. The new library design featured in this updated kit adds an 8 nucleotide (nt) unique molecular identifier (UMI) through the reverse-transcription step to mitigate potential PCR bias as well as to provide customers with additional information for transcript quantification, specifically for true variants and rare mutations. This kit includes the SMARTer RNA Unique Dual Index Kit - 24U (Takara Bio, Cat. No. 634451). SMARTer RNA Unique Dual Index Kit – 96U Set A and B (Cat. #s 634452, and 634457) are available for purchase. Together they offer up to 192 unique dual indexes for multiplexing.