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Quantitative analysis of Ig gene rearrangements

WW Wenyuan Wang
TO Tonis Org
AM Amélie Montel-Hagen
PP Peter D. Pioli
DD Dan Duan
EI Edo Israely
DM Daniel Malkin
TS Trent Su
JF Johanna Flach
SK Siavash K. Kurdistani
RS Robert H. Schiestl
HM Hanna K. A. Mikkola
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Genomic DNA was extracted from sorted B-cell populations (pro-B for heavy chain and pre-B for light chain) with the genomic DNA extraction kit (QIAGEN). Quantitative analysis of VH-J558 and VH-7183 heavy chain and κ and λ light chain rearrangements were performed by Q-PCR using published primers61. Q-PCR was performed as stated above. Samples were normalized to Actb. Rearrangement frequencies were calculated as 2ΔCt with ΔCt=CtActb–CtIg.

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