Genetic mapping and imprinting studies
A 469 bp product corresponding to the 3' end of Ndnl2 was generated from mouse genomic DNA (C57BL/J or SPRET/Ei) using oligonucleotide primers Ndnl2-7F (5'-TGGAAACCAGCAAGATGAAA) and Ndnl2-8R (5'-AGCTACCCTGTTTCTTTATCGTC). DNA samples were sequenced on both strands using the LiCor automated sequencer. DNA amplification products were digested with MspI to produce a 469 bp undigested product in C57BL/6J and 180 bp and 289 bp digested products in DNA from M. spretus. Genetic mapping was performed on the Jackson Laboratories BSS backcross. Cell lines used for human imprinting analysis were PWS lymphoblast GM09024B and AS lymphoblast GM11515 (both from the NIGMS Human Genetic Mutant Cell Repository), PWS fibroblast 1889 from the University of Miami Brain and Tissue Bank for Developmental Disorders and AS fibroblast KAT graciously provided by Dr. A. Beaudet (Baylor College of Medicine).