As a lab within the Cooperative Center of Excellence in Hematology (CCEH) at Fred Hutch, the Cell and Molecular Services and Analysis lab provides VNTR (Variable Number Tandem Repeat), and LAM-PCR (Linear Amplification Mediated PCR) and other molecular technology to analyze genotypes. Services were established initially to assist hematological research, but the facility also offers our services to scientists in other areas of research. We also provide products, protocols, and services for cell and molecular analyses of the Major Histocompatibility Complex (MHC).
Bone marrow stromal lines: Several cell lines have been developed in the Torok-Storb lab from animal bone marrow stroma. These lines have been used to support hematopoiesis in vitro, and to study the cellular interactions involved in this development.
Purified hematopoietic cell populations: A state-of the-art large-scale cell processing core, makes it possible to generate immuno-magnetically enriched populations of animal hematopoietic cells, including CD34+, CD3+ and CD14+ populations. Additional populations may become available, so please inquire.
MHC Class I Typing: DLA-88 is the most polymorphic Class I locus and is the primary focus of DLA class I typing.
Typical analysis of DLA-88 includes amplification of a 1.4 kb region of chromosome 12 which contains the polymorphic exons, 2 and 3 and flanking introns of DLA-88. The PCR product is amplified with a high-fidelity DNA polymerase, cloned, and sequenced.
For descriptions of most known alleles, to date, see Kennedy et al., (2001) Animal Genetics 32:193-199; Hardt et al. (2006) Tissue Antigens 67:163-165; and Venkataraman et al. (2007) Tissue Antigens 70:53-57
We can provide primers and protocols if you wish to do the DLA typing yourself.