Can nuclei from frozen samples be used for V(D)J enrichment?

2026-06-23

Answer:
No. Nuclei are not suitable for V(D)J enrichment.

Reasons:

  • Loss of cytoplasmic mRNA:
    Nuclei isolation removes cytoplasm, where most mature mRNA (including V(D)J transcripts) resides. 
  • Low transcript abundance:
    V(D)J transcripts are already relatively low in abundance, making them even harder to detect after cytoplasmic loss. 
  • High background RNA:
    Nuclei preparation may release cytoplasmic RNA into the environment, increasing ambient RNA noise and interfering with accurate signal detection and V(D)J assembly. 

Recommendation: Use fresh or properly cryopreserved whole-cell suspensions.

 

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