What is the size of adapters added to the SMARTer® Human BCR IgG IgM H/K/L library?

Total size of adapter-dimer in theSMARTer Human BCR IgG IgM H/K/L library is 159bp - as shown in the SMARTer® Human BCR IgG IgM H/K/L Profiling Kit User Manual -Figure 5. SMARTer Human BCR IgG IgM H/K/L Profiling library structure. First 19 nt from Read2 could be trimmed off if UMI analysis is not performed.Immune profiling [...]

What brand of beads is compatible with the SMARTer® Human BCR IgG IgM H/K/L Profiling Kit workflow?

NucleoMag NGS Clean-up and Size Select (available from Takara Bio; 5-ml size: Cat. No. 744970.5; 50-ml size: Cat. No. 744970.50; 500-ml size: Cat. No. 744970.500) or AMPure XP PCR purification kit (Beckman Coulter; 5-ml size: Cat. No. A63880; 60-ml size: Cat. No. A63881) is an appropriate substitute, using the same bead:sample ratios as recommended for the NucleoMag NGS Clean-up and Size [...]

What read length is recommended for the SMARTer® Human BCR IgG IgM H/K/L libraries?

For read through the entire V(D)J reagion use 600-cycle MiSeq Reagent Kit v3 with 2 x 300 base pair reads. To cover the entire CDR3 region and UMI use Read1-200 bp and Read2- 50bp.Immune profiling [...]

Is the SMARTer Human TCR a/b Profiling Kit v2 compatible with species other than Human?

SMARTer Human TCR a/b Profiling Kit v2 has been validated for TCR using RNA from rhesus macaque PBMC follwing the standard protocol , as described in the Technical Note - Highly reproducible TCR profiling using RNA from rhesus macaque PBMC.Immune profiling [...]

Can the Cogent NGS Immune Profiler Software analyze only Read1 sequencing data?

No. Read 2 is essential for the Cogent NGS Immune Profiler Software analysis, since it carries UMIs as shown in the SMARTer Human TCR a/b Profiling Kit v2 User Manual - Figure 4. SMARTer human TCR profiling v2 library structure. Cogent NGS Immune Profiler Software requires the presence of UMIs for the analysis.Immune profiling [...]