Abstract
This protocol is used for the detection of immunoglobulin heavy (H) chain rearrangements. This PCR-based assay enables detection of DH-JH recombination in cultured hematopoietic cells (Schlissel et al., 1991; Satoh et al., 2013) [e.g. ES-derived cells (Satoh et al., 2013)].
Materials and Reagents
Equipment
Procedure
Results
Figure 1.DNA PCR assays of germline (GL) or DH-JH rearranged Igh chain (DJ) genes were performed with mouse splenocytes. A PCR experiment using a primer DHL(5’) and J3(3’) can detect three types of DH-JH rearrangement (J1, J2, and J3) (Schlissel et al., 1991). All of three bands are present with successful DH-JH rearrangement. However, a J1 band is sometimes undetected in the ethidium bromide-based DNA-band visualization when the amount of template DNA is very small. The size marker was loaded in the left lane.
Acknowledgments
This protocol was adapted from a previously published paper by Schlissel et al. (1991). The representative data shown in the protocol was adapted from Satoh et al. (2013).
References
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