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. Author manuscript; available in PMC: 2013 Nov 20.
Published in final edited form as: J Immunol Methods. 2010 Jan 28;354(0):10.1016/j.jim.2010.01.007. doi: 10.1016/j.jim.2010.01.007

Figure 3. CD B cells exhibit non-biased usage of several VH-family genes.

Figure 3

Non-adherent BL/6 BM cells were cultured (1°-IL-7, 2°-IL-7) to generate CD B cells. Cells were harvested and labeled with mAbs to B220, IgM and IgD. Pro-/pre-B cells were sorted from BM (B6) or 2°CD B cells (CD) and genomic DNA was extracted from 106 cells using proteinase K digestion and phenol-chloroform extraction. Whole BM (BM) gDNA sample was used as positive control for detection. ∼20 ng of gDNA was amplified by q-rtPCR using VH1-, VH2- or VH9-family specific 5` primer with an intronic JH-1 specific 3` primer. CD14-specific PCR reaction was used to normalize input quantities of gDNA. Values are represented as the percent of CD14 signal. Experiment was performed on sorted samples (n=3 each) of BL/6 BM and 2° CD pro-/pre-B cells.