Question: Why are there two target enrichment steps in the Single Cell V(D)J workflow?
Answer: The two enrichment steps in the Single Cell V(D)J workflow improve specificity by utilizing a hemi-nested PCR strategy. The first PCR step utilizes primers targeting the P5 and Read 1 sequence (forward primer) and a distal position of the TCR or Ig constant regions (reverse primer), and the second PCR step targets the P5 and partial R1 sequence (forward primer) and a proximal position in the constant regions (reverse primer).
For additional information on the assay scheme and primer sequences, please see the appendix section of the Single Cell V(D)J User Guide:https://support.10xgenomics.com/single-cell-vdj/library-prep/doc/user-guide-selecting-the-correct-single-cell-immune-profiling-user-guide
Products: Single Cell Immune Profiling