Construction and characterization of partial digest DNA libraries made from flow-sorted human chromosome 16.
In this report, we present the techniques used for the construction of chromosome-specific partial digest libraries from flow-sorted chromosomes and the characterization of two such libraries from human chromosome 16. These libraries were constructed to provide materials for use in the development of a high-resolution physical map of human chromosome 16, and as part of a distributive effort on the National Laboratory Gene Library Project. Libraries with 20-fold coverage were made in Charon-40 (LA16NL03) and in sCos-1 (LA16NC02) after chromosome 16 was sorted from a mouse-human monochromosomal hybrid cell line containing a single homologue of human chromosome 16. Both libraries are approximately 90% enriched for human chromosome 16, have low nonrecombinant backgrounds, and are highly representative for human chromosome-16 sequences. The cosmid library in particular has provided a valuable resource for the isolation of coding sequences, and in the ongoing development of a physical map of human chromosome 16.
Longmire
J L
JL
Life Sciences Division and Center for Human Genome Studies, Los Alamos National Laboratory, NM 87545.
Brown
N C
NC
Meincke
L J
LJ
Campbell
M L
ML
Albright
K L
KL
Fawcett
J J
JJ
Campbell
E W
EW
Moyzis
R K
RK
Hildebrand
C E
CE
Evans
G A
GA
D000818:Animals; D002885:Chromosomes, Human, Pair 16; D003001:Cloning, Molecular; D003360:Cosmids; D004247:DNA / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific; D005434:Flow Cytometry; D015723:Gene Library; D006801:Humans; D006822:Hybrid Cells; D051379:Mice; D010766:Phosphorylation
D004247:DNA; D015252:Deoxyribonucleases, Type II Site-Specific; C024405:GATC-specific type II deoxyribonucleases