TY - JOUR
T1 - Saturation of human chromosome 3 with unique sequence hybridization probes
AU - Smith, David I.
AU - Mangrulker, Rajesh
AU - Geist, Rose
AU - Gilbert, Jeffrey
AU - Kinsman, Kirstein
AU - Drabkin, Harry
AU - Golembieski, William
N1 - Funding Information:
This work was supported by March dation Grant 5-645, by Grant lN-162 Society, and by the Molecular Biology versity.
PY - 1989/5
Y1 - 1989/5
N2 - We have generated chromosome 3-specific recombinant libraries in both λ and cosmid cloning vectors starting with somatic cell hybrids (hamster/human) containing either an intact chromosome 3 or a chromosome 3 with an interstitial deletion removing 75% of long-arm sequences. The libraries contained between 2 × 105 and 5 × 106 independent recombinants. Approximately 2% of the recombinants in these libraries contain inserts of human DNA. These were identified by hybridizing the recombinants to radioactively labeled total human DNA. Over 2500 recombinants containing human DNA were isolated from these various libraries and DNA was prepared from each of them. This represents 80,000 kb of cloned chromosome 3 sequences. One-third of the DNAs were digested with EcoRI or HindIII, and fragments free of repetitive sequences were radioactively labeled using random hexanucleotide primers and tested as unique sequence hybridization probes. Over 6500 of the fragments were tested and of these 758 were unique sequence probes with minimal or no background hybridization. Their hybridization only to chromosome 3 was verified. These probes, which were derived from 452 independent recombinants, should provide an effective saturation of human chromosome 3.
AB - We have generated chromosome 3-specific recombinant libraries in both λ and cosmid cloning vectors starting with somatic cell hybrids (hamster/human) containing either an intact chromosome 3 or a chromosome 3 with an interstitial deletion removing 75% of long-arm sequences. The libraries contained between 2 × 105 and 5 × 106 independent recombinants. Approximately 2% of the recombinants in these libraries contain inserts of human DNA. These were identified by hybridizing the recombinants to radioactively labeled total human DNA. Over 2500 recombinants containing human DNA were isolated from these various libraries and DNA was prepared from each of them. This represents 80,000 kb of cloned chromosome 3 sequences. One-third of the DNAs were digested with EcoRI or HindIII, and fragments free of repetitive sequences were radioactively labeled using random hexanucleotide primers and tested as unique sequence hybridization probes. Over 6500 of the fragments were tested and of these 758 were unique sequence probes with minimal or no background hybridization. Their hybridization only to chromosome 3 was verified. These probes, which were derived from 452 independent recombinants, should provide an effective saturation of human chromosome 3.
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U2 - 10.1016/0888-7543(89)90268-1
DO - 10.1016/0888-7543(89)90268-1
M3 - Article
C2 - 2744758
AN - SCOPUS:0024670871
SN - 0888-7543
VL - 4
SP - 453
EP - 459
JO - Genomics
JF - Genomics
IS - 4
ER -