Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I

Rigby, Peter; Dieckmann, Marianne; Rhodes, C. Harker; Berg, Paul

doi:10.1016/0022-2836(77)90052-3

Cited by 11,411 publications

(3,280 citation statements)

References 35 publications

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“…A 0.96 kb EcoRI fragment of this partial clone was nicktranslated with (o~-nP)-labeled dATP [10] and used for library screens, Northern and Southern blots. The probe represented the 3'-region downstream of the internal EcoRI site (fig.l).…”

Section: Edna and Genornic Cloningmentioning

confidence: 99%

A developmentally regulated gene product from Dictyostelium discoideum shows high homology to human α‐L‐fucosidase

et al. 1989

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A cDNA library of poly(A+)-RNA has been prepared from membrane-bound polysomes of Dictyostelium discoideum and screened for clones hybridizing to mRNA species that encode developmentally regulated proteins. The clone investigated in this paper recognizes a 1.8 kb transcript that accumulates strongly between the growth phase and aggregation stage. Stimulation of cells with pulses of CAMP enhances the accumulation. The amino acid sequence derived from a complete cDNA and from a genomic clone displays extensive sequence identity to human liver a-L-fucosidase. The D. discoideum DNA sequence encodes a 50.5 kDa polypeptide with a hydrophobic signal peptide at the N-terminus. Antibodies against a synthetic peptide corresponding to amino acids 262-275 of the deduced protein sequence recognize a developmentally regulated 50 kDa protein in D. discoideum that is recovered in the particulate fraction.

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Section: Edna and Genornic Cloningmentioning

confidence: 99%

A developmentally regulated gene product from Dictyostelium discoideum shows high homology to human α‐L‐fucosidase

et al. 1989

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“…The probes VIII, XIII and cVII were 2.5, 0.46 and 2.0 kb in length, respectively. They were radiolabeled by nick translation (Rigby et al, 1977), and were denatured by heating before hybridisation.…”

Section: Subjectsmentioning

confidence: 99%

RFLPs of factor IX gene in Japanese haemophilia B families and gene deletion in two high-responder-inhibitor patients

et al. 1987

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SummaryThe factor IX genes in four Japanese families with haemophilia B were analysed for the restriction fragment length polymorphisms (RFLPs) of TaqI, Xmnl and DdeI, using subcloned intragenic DNA fragments as probes (probes VIII and XIII). The factor IX genes in 12 patients with haemophilia B and three high-responder-inhibitor cases showed no size difference using a cDNA probe (cVII) when restricted by TaqI, EcoRI and HindlII. Complete gene deletions were observed in two other highresponder-inhibitor cases.

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“…Filters were then washed in 3 x SSC and dried for 2-3 h at 80°C. Prehybridization (2 h) and hybridization (12-18 h) were performed in the medium described in [13] with 'nick translated' DNA probes [14]. Filters were washed at 65°C in 0.1 x SSC/O.l% SDS.…”

Section: Dot-spot and Filter Hybridizationmentioning

confidence: 99%

Rapid identification of Trypanosoma cruzi isolates by ‘dot‐spot’ hybridization

Sánchez¹,

Madrid²,

Engel

et al. 1984

FEBS Letters

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The presence of isolate-specific Trypanosoma cruz~ minicircles has been shown in the kinetoplast DNA of this parasite. This led to the rapid identification of isolates and clones of trypanosomes by means of 'dot-spot' hybridizations with molecularly cloned minicircle probes. Unexpectedly, whole kDNAs were also suitable as probes for this purpose, provided that filters were washed under stringent conditions. This was attributed to the presence of the above-mentioned isolate-specific minicircle sequences. The fact that parasites could be directly spotted onto nitrocellulose filters simplified the rapid routine screening of a large number of samples.

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Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I

Cited by 11,411 publications

References 35 publications

A developmentally regulated gene product from Dictyostelium discoideum shows high homology to human α‐L‐fucosidase

A developmentally regulated gene product from Dictyostelium discoideum shows high homology to human α‐L‐fucosidase

RFLPs of factor IX gene in Japanese haemophilia B families and gene deletion in two high-responder-inhibitor patients

Rapid identification of Trypanosoma cruzi isolates by ‘dot‐spot’ hybridization

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