Construction of Human Chromosome-specific DNA Libraries from Flow-sorted Chromosomes

Deaven, L.L.; Dilla, M. A. Van; Bartholdi, Marty F.; Carrano, A.V.; Cram, L.S.; Fuscoe, J.C.; Gray, J. W.; Hildebrand, C.E.; Moyzis, Robert K.; Perlman, J.

doi:10.1101/sqb.1986.051.01.019

Cited by 99 publications

(35 citation statements)

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“…Cosmid clones were obtained by PCR screening of 110 plates of the LA08Nco1 flow-sorted chromosome 8-specific cosmid library (Deaven et al 1986) using a plate-pool strategy. PCR reactions were carried out in a 20-µl volume consisting of the following: 100 ng of DNA, ‫ן1‬ PCR buffer with 1.5 mM MgCl 2 (PE Applied Biosystems), 0.1 mM dNTPs (Pharmacia), 1 µM of each primer, and 1 unit of AmpliTaq DNA polymerase (PE Applied Biosystems).…”

Section: Methods Pcr-based Library Screeningmentioning

confidence: 99%

“…Figure 1B shows the names and approximate extent of the selected YAC clones that encompass the deletion interval, of which YAC clones 946_c_9 and 932_e_9 had been used previously to develop a long-range restriction map of the deletion interval depicted in Figure 1C ( Bookstein et al 1994). To prepare a physical contig, PCR was used to screen human genomic BAC (Shizuya et al 1992) and P1 (Pierce et al 1992) pooled DNA libraries, as well as a cosmid library prepared from flowsorted chromosome 8 (Deaven et al 1986), using the primer pairs listed in Table 1. A total of 39 sequence tagged sites (STSs) were used for library screening: 20 are published, 2 were a gift from Dr. R. Bookstein (unpubl.…”

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confidence: 99%

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High-Resolution Physical Map and Transcript Identification of a Prostate Cancer Deletion Interval on 8p22

Arbieva¹,

Banerjee²,

Kim³

et al. 2000

Genome Res.

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A genomic interval of ∼1-1.5 Mb centered at the MSR marker on 8p22 has emerged as a possible site for a tumor suppressor gene, based on high rates of allele loss and the presence of a homozygous deletion found in metastatic prostate cancer. The objective of this study was to prepare a bacterial contig of this interval, integrate the contig with radiation hybrid (RH) databases, and use these resources to identify transcription units that might represent the candidate tumor suppressor genes. Here we present a complete bacterial contig across the interval, which was assembled using 22 published and 17 newly originated STSs. The physical map provides twofold or greater coverage over much of the interval, including 17 BACs, 15 P1s, 2 cosmids, and 1 PAC clone. The position of the selected markers across the interval in relation to the other markers on the larger chromosomal scale was confirmed by RH mapping using the Stanford G3 RH panel. Transcribed units within the deletion region were identified by exon amplification, searching of the Human Transcript Map, placement of unmapped expressed sequence tags (ESTs) from the Radiation Hybrid Database (RHdb), and from other published sources, resulting in the isolation of six unique expressed sequences. The transcript map of the deletion interval now includes two known genes (MSR and N33) and six novel ESTs.

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Section: Methods Pcr-based Library Screeningmentioning

confidence: 99%

mentioning

confidence: 99%

High-Resolution Physical Map and Transcript Identification of a Prostate Cancer Deletion Interval on 8p22

Arbieva¹,

Banerjee²,

Kim³

et al. 2000

Genome Res.

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“…Human chromosomes were isolated and sorted directly on nitrocellulose discs as previously described [14,15] and hybridized to labelled Gp IX cDNA.…”

Section: Chromosomal Localizationmentioning

confidence: 99%

Human platelet glycoprotein IX

1990

FEBS Letters

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Overlapping cDNAs encoding human platelet glycoprotein (Gp)IX were cloned from a human erythroleukemia cell λ gtll library. The possibly ‘full‐length’ cDNA of 896 base pairs (bp) includes an open reading frame (528 bp), both 5' (222 bp) and 3' (146 bp) noncoding regions, and a poly(A) tail. Translation predicts a signal peptide of 16 amino acids and a mature protein of 160 amino acids that includes a 24 amino acid leucinerich glycoprotein (LRG) segment. Southern blot analysis suggests the presence of a single copy of the Gp IX gene, and hybridization of Gp IX cDNA to sorted human chromosomes localizes the Gp IX gene to chromosome 3.

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“…The majority of DNA probes (series pFS) were isolated from a flow-sorted human chromosome 7library in phage Charon 21A (LA07NS01) (Deaven et al, 1986). Plaques were screened according to Benton and Davis (1977) with either total genomic human or total genomic hamster DNA labeled with 32 P by random primed DNA labeling.…”

Section: Dna Probesmentioning

confidence: 99%

A somatic cell hybrid panel and DNA probes for physical mapping of human chromosome 7p

et al. 1991

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To identify by reverse 1enetics genes on the short arm of human chromosome 7 ex~ted to ~ involved b:a the regulation of human craniofacial and limb development, we have set up a human mouse somatic cell hybrid panel that divides 7p into 9 fragments. The breakpoints are defined by deletions or translocations involving one chromosome 7 in the cells of the human cell fusion partners. Particularly densely covered with these cytogenetic anchor points is the proximal area of 7p within and around 7p13. The number of cytogenetic mapping points within proximal 7p could be increased by four, using two diploid human celllines with small interstitial deletions in this region for dosage studies. We used Southern blots oftbis panel to assign to 7 q or subregions of 7p more than 300 arbitrary DN A probes or genes that provide reference points for physical mapping of 7p. Three reciprocal translocations with one of the breakpoints in 7pl3 mark tbe location of a gene involved in Greig cephalopolysyndactyly syndrome. To deftne an area in wbich we could identify candidatel fortbis developmental gene, we established a macrorestriction map using probe& ftanking the putative gene region. The Greig translocations were found tobe located within a 630-kb Notl restriction fragment.

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Construction of Human Chromosome-specific DNA Libraries from Flow-sorted Chromosomes

Cited by 99 publications

References 0 publications

High-Resolution Physical Map and Transcript Identification of a Prostate Cancer Deletion Interval on 8p22

High-Resolution Physical Map and Transcript Identification of a Prostate Cancer Deletion Interval on 8p22

Human platelet glycoprotein IX

A somatic cell hybrid panel and DNA probes for physical mapping of human chromosome 7p

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