Why are the 5’-ends of PCR Primer IIA and 3’ SMART-Seq CSD Prime IIA are blocked?

The 5’-ends of the PCR Primer IIA and 3’ SMART-Seq CSD Prime IIA, included in SMART-Seq mRNA Kit, are blocked to prevent ligation of sequencing adapters to DNA fragments carrying the SMART sequences, thus preventing the SMART sequences from entering the sequencing reads. Ultra-low input NGS-RNA [...]

What is the advantage of using the LNA (lock nucleic acid) template-switching oligo for the SMART(er) cDNA synthesis?

LNA template-switching oligo has a higher has a higher affinity to non-template nucleotides added by the MMLV-derived SMARTScribe reverse transcriptase at the 5’-end of the RNA template, thus enabling a more efficient template switch and, consequently a higher number of identified genes, including genes expressed at lower levels. Please, see the Tech Note - Improved Chemistry for Single-Cell [...]

What is the advantage of using SeqAmp DNA polymerase in SMART(er) cDNA synthesis for NGS?

SeqAmp DNA polymerase is characterized by a robust amplification of GC-rich templates, thus ensuring a better representation of GC-rich sequences in the cDNA [...]

What is the advantage of using SMARTScribe Reverse Transcriptase for the SMART-Seq mRNA cDNA synthesis?

SMARTScribe RT is a modified Moloney Murine Leukemia Virus Reverse Transcriptase that generates long, full-length cDNA (up to 14.7 kb) thus ensuring a true representation of the original mRNA pool in the SMART(er) Ultra Low cDNA library. Please, also see Application Note SMARTScribe Reverse Transcriptase—the RT of choice for SMART and SMARTer Kits.Ultra-low input NGS-RNA [...]

Does the SMART-Seq mRNA cDNA synthesis require capped mRNA?

No, SMART-Seq mRNA cDNA synthesis is cap-independent, ensuring a broad representation of the original mRNA pool in the SMART-Seq cDNA library.Ultra-low input NGS-RNA [...]