Literature DB >> 3218734

A quantitative method for analyzing specific DNA sequences directly from whole cells.

P McIntyre1, G R Stark.   

Abstract

A quick, accurate assay for specific DNA sequences is described in which whole cells are treated with 0.4 M sodium hydroxide at 80 degrees C. DNA is relatively resistant to alkaline hydrolysis, whereas proteins and RNA are degraded rapidly. The DNA in NaOH is then transferred through a slot directly onto a nylon membrane and hybridized with a probe. Since the procedure is so simple, many samples can be analyzed in a short time. A single-copy gene can be detected in as few as 1000 cells and, since the DNA from 10(5) cells can be loaded through a single slot, the sensitivity is sufficient to detect one specific DNA sequence per 100 cells. Accurate quantitative analysis can be achieved by normalizing the amount of DNA available for hybridization in each slot, using a probe derived from total DNA.

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Year:  1988        PMID： 3218734     DOI： 10.1016/0003-2697(88)90537-4

Source DB:  PubMed          Journal:  Anal Biochem        ISSN： 0003-2697            Impact factor:   3.365


  10 in total

1.  DNA amplification is rare in normal human cells.

Authors:  J A Wright; H S Smith; F M Watt; M C Hancock; D L Hudson; G R Stark
Journal:  Proc Natl Acad Sci U S A       Date:  1990-03       Impact factor: 11.205

2.  Evolution and stability of chromosomal DNA coamplified with the CAD gene.

Authors:  I Saito; R Groves; E Giulotto; M Rolfe; G R Stark
Journal:  Mol Cell Biol       Date:  1989-06       Impact factor: 4.272

3.  A strategy for quickly identifying all unique two-hybrid or library plasmids within a pool of yeast transformants.

Authors:  T E Patterson; G R Stark; S Sazer
Journal:  Nucleic Acids Res       Date:  1995-10-25       Impact factor: 16.971

4.  DNA hybridisation of routinely processed tissue for detecting HPV DNA in anal squamous cell carcinomas over 40 years.

Authors:  J H Scholefield; P McIntyre; J G Palmer; P J Coates; N A Shepherd; J M Northover
Journal:  J Clin Pathol       Date:  1990-02       Impact factor: 3.411

5.  ERBB2 amplification in breast cancer analyzed by fluorescence in situ hybridization.

Authors:  O P Kallioniemi; A Kallioniemi; W Kurisu; A Thor; L C Chen; H S Smith; F M Waldman; D Pinkel; J W Gray
Journal:  Proc Natl Acad Sci U S A       Date:  1992-06-15       Impact factor: 11.205

6.  Anal human papillomavirus infection: a comparative study of cytology, colposcopy and DNA hybridisation as methods of detection.

Authors:  C Sonnex; J H Scholefield; G Kocjan; G Kelly; C Whatrup; A Mindel; J M Northover
Journal:  Genitourin Med       Date:  1991-02

7.  Interleukin 1 induction of the c-jun promoter.

Authors:  K Muegge; M Vila; G L Gusella; T Musso; P Herrlich; B Stein; S K Durum
Journal:  Proc Natl Acad Sci U S A       Date:  1993-08-01       Impact factor: 11.205

8.  Simian virus 40 large tumor antigen alone or two cooperating oncogenes convert REF52 cells to a state permissive for gene amplification.

Authors:  M E Perry; M Commane; G R Stark
Journal:  Proc Natl Acad Sci U S A       Date:  1992-09-01       Impact factor: 11.205

9.  Kappa B site-dependent activation of the interleukin-2 receptor alpha-chain gene promoter by human c-Rel.

Authors:  T H Tan; G P Huang; A Sica; P Ghosh; H A Young; D L Longo; N R Rice
Journal:  Mol Cell Biol       Date:  1992-09       Impact factor: 4.272

10.  A steroid hormone response unit in the late leader of the noncoding control region of the human polyomavirus BK confers enhanced host cell permissivity.

Authors:  U Moens; N Subramaniam; B Johansen; T Johansen; T Traavik
Journal:  J Virol       Date:  1994-04       Impact factor: 5.103
  10 in total

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