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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