Resolution of a Secondary Structure in an Unknown mRNA 5’ End

  • M. Bacher
  • P. Hofmann
  • D. Gemsa
Chapter

Abstract

Obtaining the complete cDNA is of critical importance for gene structure and expression studies. cDNA clones derived from libraries frequently lack the full-length 5’end of the sense strand. The RACE (rapid amplification of cDNA ends) PCR technique first described by Frohman et al (1988) has proven to be a versatile and fast method for obtaining the complete 5’end of cDNAs.

Keywords

Glycerol DMSO Agarose Bromide Influenza 

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References

  1. Chomczynsky P and Sacchi N. Single step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 1987, 162:156–159Google Scholar
  2. Gong JH, Sprenger H, Hinder F, Bender A, Schmidt A, Horch S, Nain M, and Gemsa D. Influenza a virus infection of macrophages: enhanced tumor necrosis factor-a (TNF-a) gene expression and lipopolysaccharide-triggered TNF-a release. J Immunol 1991, 147:3507–3513PubMedGoogle Scholar
  3. Klenk HD and Rott R. On the structure of the influenza virus envelope. Virology 1972, 47:579–588PubMedCrossRefGoogle Scholar
  4. Nain M, Hinder F, Gong JH, Schmidt A, Bender A, Sprenger H, and Gemsa D. Tumor necrosis-a production of influenza A virus-infected macrophages and potentiating effect of lipopolysaccharides. J Immunol 1990, 145:1921–1929PubMedGoogle Scholar
  5. Ruano G, Pagliaro EM, Schwartz TR, Lamy K, Messina D, Gaensslen RE and Lee HC. Heat-soaked PCR: an efficient method for DNA amplification with application to forensic analysis. Biotechniques 1992, 13:266–274PubMedGoogle Scholar
  6. Shen WH and Hohn B. DMSO improves PCR amplification of DNA with complex secondary structure. TIG 1992, 8:227CrossRefGoogle Scholar
  7. Smith KT, Long CM, Bowman B, and Manos M. Using cosolvents to enhance PCR amplification. Amplifications: A Forum for PCR Users 1990, 5:16–17Google Scholar

Copyright information

© Springer Science+Business Media New York 1994

Authors and Affiliations

  • M. Bacher
    • 1
  • P. Hofmann
    • 1
  • D. Gemsa
    • 1
  1. 1.Institute of ImmunologyPhilipps UniversityMarburgGermany

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