Can I amplify BCR IgG/IgM/IgK/IgL chains in one reaction?

For optimal balance between the different chains, we recommend amplifying them separately. If multiplex PCR is preferred, in order to amplify all four chains at the same time, we recommend combining only heavy chain primers (G+M) and light chain primers (K+L) strting with PCR1- see the SMARTer® Human BCR IgG IgM H/K/L Profiling Kit User Manual -Appendix A: Guidelines for Multiplexing Heavy [...]

What is the number of maximal theoretical UMI combinations in the SMARTer Human BCR IgG IgM H/K/L sequencing reads?

The number of maximal theoretical UMI combinations is 4^12=16,777,216. Immune profiling [...]

What is the nature of the UMIs incoporated into the SMARTer® Human BCR IgG IgM H/K/L library?

UMIs incoporated into the SMARTer® Human BCR IgG IgM H/K/L library represent a random 12 nt sequence: NNN,NNN,NNN,NNN, (N=G,T,A,C)Immune profiling [...]

What is the advantage of incoporating Unique Molecular Identifiers (UMI)?

Incorporated Unique Molecular Identifiers (UMI) eliminate PCR and sequencing errors, reduce the number of duplicates and ensure a greater confidence in clonotype identification, allowing a correct clonotype call at a lower sequencing depth, than non UMI pipeline.Immune profiling [...]

What software should I use for the analysis of the Human BCR IgG IgM H/K/L sequencing reads generated using the SMARTer® Human BCR IgG IgM H/K/L Profiling Kit?

For the analysis of the Human BCR IgG IgM H/K/L sequencing reads generated using the SMARTer® Human BCR IgG IgM H/K/L Profiling Kit we recommend free-of charge Cogent NGS Immune Profiler Software.Immune profiling [...]