Acidithiobacillus ferrooxidans check for this species in other resources , co-transcription, polycistronic mRNA"/>
search
for
 About Bioline  All Journals  Testimonials  Membership  News  Donations


Electronic Journal of Biotechnology
Universidad Católica de Valparaíso
ISSN: 0717-3458
Vol. 3, No. 3, 2000, pp. 213-216
Bioline Code: ej00022
Full paper language: English
Document type: Short Communication
Document available free of charge

Electronic Journal of Biotechnology, Vol. 3, No. 3, 2000, pp. 213-216

 en An RT-PCR artifact in the characterization of bacterial operons
Guacucano, Maritza; Levican, Gloria; Holmes, David S. & Jedlicki, Eugenia

Abstract

During the course of experiments involving RT-PCR to investigate the co-transcription of the pet and res operons of the bacterium Acidithiobacillus ferrooxidans check for this species in other resources we observed that the mRNA of certain gene pairs can be amplified by RT-PCR in the absence of added RT-primer. We provide experimental evidence that eliminates, as explanations for the observed results, the possible global contamination of the RNA preparation by genomic DNA or by contaminating single stranded DNA or RNA fragments. We suggest that the results can be explained by self-priming of the RNA perhaps as a result of secondary structure configurations that provide a suitable 3'-terminus to prime the reverse transcriptase. This type of artifact can be recognized by carrying out a control in which exogenously added RT-primers are left out of the reaction. Such a control is essential if RT-PCR is to be used to investigate and correctly identify operons. RT-PCR is also an important tool for the amplification of eukaryotic mRNAs and our observations may also be applicable to these systems.

Keywords
Acidithiobacillus ferrooxidans check for this species in other resources , co-transcription, polycistronic mRNA

 
© 2000 by Universidad Católica de Valparaíso -- Chile
Alternative site location: http://www.ejbiotechnology.info

Home Faq Resources Email Bioline
© Bioline International, 1989 - 2017, Site last up-dated on 16-Oct-2017.
Site created and maintained by the Reference Center on Environmental Information, CRIA, Brazil
System hosted by the Internet Data Center of Rede Nacional de Ensino e Pesquisa, RNP, Brazil