Two-dimensional gel analysis of complex DNA families: methodology and apparatus.
We describe a reproducible protocol for the analysis of individual members of complex mammalian gene families by gel fractionation in two dimensions within a specially designed, easily built electrophoretic apparatus. We have used this protocol to resolve the family of mouse H-2 class I genes, with approximately 30 members, as well as two different families of endogenous retroviral-like sequences, each of which has approximately 180 members dispersed throughout the genome. The results demonstrate the feasibility of using this protocol for rapid, whole genome analysis of individual animals and cell lines. Two-dimensional DNA analysis of highly repeated retroviral-like DNA families could be applied to genetic mapping and cloning experiments as well as to obtaining whole genome fingerprints in the analysis of somatic cell hybrid lines that contain a subset of chromosomes from the genome of interest.
Sheppard
R D
RD
Department of Molecular Biology, Princeton University, New Jersey 08544.
Montagutelli
X
X
Jean
W C
WC
Tsai
J Y
JY
Rose
A
A
Guénet
J L
JL
Cole
M D
MD
Silver
L M
LM
D004262:DNA Restriction Enzymes; D016521:Electrophoresis, Gel, Pulsed-Field; D015180:Electrophoresis, Gel, Two-Dimensional / Q000295:instrumentation / Q000379:methods*; D005821:Genetic Techniques; D005810:Multigene Family / Q000235:genetics*; D015203:Reproducibility of Results
D004262:DNA Restriction Enzymes