Dear Josef,
Boston Probes (now part of Applied Biosystems) has developed PNA probes
and application for microbiology using rRNA as target. Recently, we have
described a fluorescence in situ hybridization method (PNA FISH) for both
gram-negative and gram-positive bacteria as well as yeast following
fixation with paraformaldehyde (Perry-O'Keefe et al., Journal of
Microbiological Methods 47 (2001) 281-292). This generic solution-phase
PNA FISH method raises the possibility of simultaneous detection and
identification of different species in a single assay (multiplex) using
species-specific PNA probes labeled with different fluorophores and has
been applied for up to four different organisms.
PNA (Peptide nucleic acid) probes are DNA mimics obeying Watson-Crick base
pairing rules for hybridization to complementary nucleic acid targets (RNA
and DNA). Due to their uncharged, neutral backbone, PNA probes exhibit
favorable hybridization characteristics such as high specificity, strong
affinity, and rapid kinetics resulting in improved hybridization to highly
structured targets such as rRNA. For more information please see a recent
review on PNA for microbiology (Stender et al., Journal of Microbiological
Methods 48 (2002) 1-17).
Please do not hesitate to contact me if you have any questions or need
technical advise. Also, we do have PNA FISH reagent kits available for
evaluation purposes.
Best regards
Henrik Stender
Applied Biosystems
15 DeAngelo Drive
Bedford, MA 01730
Tel: 781 280 2845
Fax: 781 280 2940
Email: Henrik.Stender@AppliedBiosystems.com
Brett F Williams
06/12/2002 03:46 PM
To: Henrik N Stender/BED/PEC@PEC
cc:
Subject: FITC-labelling of bacteria
----- Forwarded by Brett F Williams/BED/PEC on 06/12/2002 03:45 PM -----
Josef Azem <josef.azem@microbio.gu.se>
06/12/2002 03:15 AM
To: Cytometry Mailing List <cytometry@flowcyt.cyto.purdue.edu>
cc:
Subject: FITC-labelling of bacteria
Dear Flowers!
Has anyone have a protocoll for FITC-labelling of bacteria?
Thanks.
Josef
----------------------------------
Medical Microbiology & Immunology
Gothenburg University
Guldhedsgatan 10A
41346 Gothenburg/Sweden
Tel: +46 31 3424471
Fax: +46 31 826976
mail: josef.azem@microbio.gu.se
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