Fluorescent Labeled Oligonucleotide Probes

The NAMs used in
this work consisted of a 6-carboxyfluorescein-labeled 14-mer oligonucleotide
formed by an LNA, 2′OMe, and PS linkage (Eurogentec, Seraing,
Belgium). This sequence was designed to target a conserved region
of the 16S rRNA in the domain Eubacteria.^{52 (link)} The synthetic phospholipids 1,2-dioleoyl-3-trimethylammonium-propane
(DOTAP, chloride salt), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine
(DOPE, chloride salt), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (DSPE-PEG, ammonium
salt), and 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(lissamine rhodamine B sulfonyl) (Rh-PE) were purchased
from Avanti Polar Lipids (Alabaster, AL, USA). 4′,6-Diamidine-2′-phenylindole
dihydrochloride (DAPI) was purchased from Sigma-Aldrich (Lisbon, Portugal).
All other reagents were of analytical grade.

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Moreira L., Guimarães N.M., Pereira S., Santos R.S., Loureiro J.A., Pereira M.C, & Azevedo N.F. (2022). Liposome Delivery of Nucleic Acids in Bacteria: Toward In Vivo Labeling of Human Microbiota. ACS Infectious Diseases, 8(7), 1218-1230.

Publication 2022

DSPE-PEG 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(lissamine rhodamine B sulfonyl) (Rh-PE) 4′,6-Diamidine-2′-phenylindoledihydrochloride (DAPI)

16s rrna Alabaster Carboxyfluorescein Chloride salt Diamidine Dioleoyl phosphoethanolamine Dotap Dspe Eubacteria Lipids Lissamine rhodamine b Methoxy polyethylene glycol Nams Phosphoethanolamine Phospholipids Propane

Corresponding Organization : Universidade do Porto

Top 5 similar protocols

Variable analysis

independent variables

LNA, 2'OMe, and PS linkage of the 14-mer oligonucleotide

dependent variables

Targeting a conserved region of the 16S rRNA in the domain Eubacteria

control variables

6-carboxyfluorescein label on the 14-mer oligonucleotide
Synthetic phospholipids: DOTAP, DOPE, DSPE-PEG, and Rh-PE
DAPI (4',6-diamidine-2'-phenylindole dihydrochloride)

positive controls

Not specified

negative controls

Not specified

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