Improvement of the cDNA-AFLP method using fluorescent primers for transcription analysis in bacteria.
Here the cDNA-fluorescent amplified fragment length polymorphism (cDNA-FAFLP) technique, an improvement of cDNA-AFLP method, was used in order to analyse expression profiling in bacteria. The obtained results were validated by real-time PCR. This is the first report that validates the foreseen transcription pattern by cDNA-FAFLP with the application of real-time PCR in bacteria. This new protocol offers the possibility of quick and reliable analysis of transcription profiles, also avoids the problem linked with the use of radioisotopes, and allows a quick identification of genes differentially expressed in bacteria.
Decorosi
Francesca
F
Dipartimento di Biotecnologie Agrarie, Sezione di Microbiologia, Università degli Studi di Firenze, Piazzale delle Cascine 24, I-50144, Firenze, Italy.
Viti
Carlo
C
Mengoni
Alessio
A
Bazzicalupo
Marco
M
Giovannetti
Luciana
L
D001419:Bacteria / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D042521:Burkholderiaceae / Q000235:genetics / Q000378:metabolism; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism*; D005456:Fluorescent Dyes; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D017931:DNA Primers; D018076:DNA, Complementary; D005456:Fluorescent Dyes