Rapid cloning of multiple amplified nucleotide sequences from human neuroblastoma cell lines by phenol emulsion competitive DNA reassociation

Yosef Shiloh, Elise Rose, Chris Colletti-Feener, Bruce Korf, Louis M. Kunkel, Samuel A. Latt*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

17 Scopus citations

Abstract

A protocol for the rapid cloning of many DNA fragments from an amplified genomic region is described. The procedure is based on a modification of the phenol-emulsion reassociation technique (PERT) previously used to clone DNA fragments missing from a chromosomal deletion [Kunkel et al., Proc. Natl. Acad. Sci. USA 82 (1985) 4778-4782]. The procedure was used to construct recombinant libraries in the plasmid pBR322 which were highly enriched for amplified sequences from two neuroblastoma cell lines, CHP-126 and IMR-32. Many new amplified DNA fragments were isolated from these libraries, indicating that the PERT methodology should be of general use in isolating amplified DNA from other cell lines and tumors.

Original languageEnglish
Pages (from-to)53-59
Number of pages7
JournalGene
Volume51
Issue number1
DOIs
StatePublished - 1987
Externally publishedYes

Funding

FundersFunder number
National Institutes of Health
American Cancer SocietyCD-36
Eunice Kennedy Shriver National Institute of Child Health and Human DevelopmentP01HD018658

    Keywords

    • PERT methodology
    • Recombinant DNA
    • gene libraries
    • neuroectoderm tumors

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