Skip to main content
Log in

An effective elimination of false positives isolated from differential display of mRNAs

  • Departments
  • Hints and Tips
  • Published:
Molecular Biotechnology Aims and scope Submit manuscript

Abstract

A reverse Northern analysis that effectively eliminates the false positives isolated from differential display of mRNAs (DD) is demonstrated. Preparation of probe by one-step labeling in reverse transcription reaction is found to be more effective and specific as compared to the preparation of probe by random priming of reverse transcribed cDNA pool. Reverse Northern assay of DNA fragments isolated from DD prior to their cloning into plasmid, analysis of multiple fragments on single slot blot, and requirement of RNA only in small amounts as compared to conventional Northern makes the protocol quick, effective, and economic.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Liang, P. and Pardee, A. B. (1992) Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction.Science 257, 967–971.

    Article  PubMed  CAS  Google Scholar 

  2. Zhao, S., Ooi, S. L., and Pardee, A. B. (1995) New primer strategy improves precision of differential display.BioTechniques 18, 842–850.

    PubMed  CAS  Google Scholar 

  3. Tokuyama, Y. and Takeda, J. (1995) Use of33P-labeled primer increases the sensitivity and specificity of mRNA differential display.BioTechniques 18, 424, 425.

    PubMed  CAS  Google Scholar 

  4. Reeves, S. A., Rubio, M-P., and Louis, D. N. (1995) General method for PCR amplification and direct sequencing of mRNA differential display products.BioTechniques 18, 18–20.

    PubMed  CAS  Google Scholar 

  5. McClelland, M., Francoise M-D., and Welsh, J. (1995) RNA fingerprinting and differential display using arbitrarily primed PCR.Trends Genetics 11, 242–246.

    Article  CAS  Google Scholar 

  6. Callard, D., Lescure, B., and Mazzolini, L. (1994) A method for the elimination of false positives generated by the mRNA differential display technique.BioTechniques 16, 1906–1103.

    Google Scholar 

  7. Mou, L., Miller, H., Li, J., Wing, E., and Chalifour, L. (1994) Improvements to the differential display method for gene analysis.Biochem. Biophys. Res. Commun. 199, 564–569.

    Article  PubMed  CAS  Google Scholar 

  8. Chang, A. C.-M., Janosi, J., Hulsbeek, M., Jong, D. de, Jeffrey, K. J., Noble, J. R., and Reddel, R. R. (1995) A novel human cDNA highly homologous to the fish hormone stanniocalcin.Mol. Cell. Endocrinol. 112, 241–247.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wadhwa, R., Duncan, E., Kaul, S.C. et al. An effective elimination of false positives isolated from differential display of mRNAs. Mol Biotechnol 6, 213–217 (1996). https://doi.org/10.1007/BF02740775

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02740775

Index Entries

Navigation