Is FFPE RNA compatible with the SMARTer® Human BCR IgG IgM H/K/L Kit?

Since the first-strand cDNA synthesis of the SMARTer Human BCR profiling workflow is primed by the oligo(dT) primer, generating full-length cDNA in the initial step, input RNA should be of high integrity. Therefore, FFPE RNA would not be compatible with the SMARTer® Human BCR a/b Profiling Kit: 3’-end truncated RNA will not be dT primed and will not be incorporated into the final BCR library, [...]

Does SMARTer Human BCR IgG IgM H/K/L Profiling Kit require rRNA depletion prior to cDNA synthesis?

No, SMARTer Human BCR IgG IgM H/K/L Profiling Kit utilizes oligo(dT) priming for the first-strand cDNA synthesis (annealing to poly(A) mRNAs) followed by BCR-specific amplification, thus generating BCR-specific cDNA. It is advisable to use high integrity (RIN>7) input RNA for the SMARTer Human BCR IgG IgM H/K/L Profiling Kit.Immune profiling [...]

Is SMARTer Human BCR IgG IgM H/K/L Profiling Kit compatible with RNA purified from whole blood?

Yes, SMARTer Human BCR IgG IgM H/K/L Profiling Kit is compatible using no less than 100ng RNA purified from whole blood. The protocol may require adjustment.Immune profiling [...]

What are the input requirements for the SMARTer Human BCR IgG IgM H/K/L Profiling Kit?

SMARTer Human BCR IgG IgM H/K/L Profiling Kit utilizing oligo(dT) priming for the first-strand cDNA synthesis is compatible with high integrity total RNA (RIN>7) using 10 ng to 1 µg of total RNA from peripheral blood mononuclear cells (PBMCs) , 1 to 100 ng of total RNA from B cells, no less than 100ng Whole Blood RNA (the protocol may require adjustments)Immune profiling [...]

Can I load on the same lane of Illumina sequencer SMARTer® Human BCR IgG IgM H/K/L cDNA libraries together with libraries prepared by alternative methods?

We usually do not recommend loading SMARTer BCR libraries with libraries prepared by other methods, since the average size of the SMARTer BCR library is 600-800bp, while the average size of most of Illumina libraries prepared by other methods is about 300bp. It is imperative that all the libraries are pooled in equimolar quantities (with compatible indexes) – otherwise libraries with smaller [...]