Method for analyzing intermediate product of secondary rearrangement of Igk gene and kit
A gene and product technology, applied in the field of capturing and amplifying secondary rearrangement intermediate products of Igκ gene, can solve the problems of low sensitivity and specificity, no modification of downstream primers, and uncertain whether the VκJκ gene fragment is secondary rearranged by Igκ gene Eliminate problems such as gene fragments to achieve the effect of improving sensitivity and specificity
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[0045] 1. Take MRL.FAS lpr 、B6.FAS lpr Bone marrow single cell suspension of C3H mice was used as experimental material, and bone marrow B lymphocytes of different developmental stages were obtained by flow cytometry technology.
[0046] Prepare mouse bone marrow single cell suspension, and design antibody staining strategy according to the cell types and their specific surface antigens in the mouse bone marrow single cell suspension, as well as the specific surface antigens of B cells at various developmental stages, so as to sort wild type mice (C3H) and lupus erythematosus mice (B6.FAS lpr , MRL.FAS lpr ) pro-B (B220 + CD43 + IGM - IGD - ), small pre-B (B220 + CD43 - IGM - IGD - ) and immature-B (B220 + CD43 - IGM + IGD - )cell.
[0047] 2. Extraction of genomic DNA from pro-B, small pre-B and immature-B cells Extract genomic DNA from pro-B, small pre-B and immature-B cells obtained by sorting. Due to the number of cells obtained by sorting Less and subseque...
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