THE MOLECULAR PROBE :

Applications of chemistry in biomedicine.

Mirella Lozano Álvarez y

Miruna Nicoleta Nedelcu

02/02/2023
3.º ESO, F SCIENCE

WHAT IS A MOLECULAR PROBE?

Molecular probes are small pieces of DNA or RNA that are used in
molecular biology as tools to detect the presence of DNA or RNA with
similar or identical complementary sequences.

The probe binds to a single-stranded target sequence (the only strand

remaining when DNA or RNA is isolated) by a hybridization mechanism
that forms a double-stranded structure, one strand belonging to the probe
and the other to the target sequence. This association is formed because
both the probe and the target have partially or fully complementary
sequences that form complementary base pairs. This is the process:
 1. We extract DNA samples from the subjects we want to analyze and
make a molecular probe with the same nitrogenous base.
2. The DNA fibers are separated with heat or enzymes, and the probe is
then attached to one of the fibers.
3. Later a wash will be performed to remove the DNA fiber. If the strand
then fluoresces, it means that the DNA matches and the subjects are
related.
4. If nothing is seen it means that the DNA does not match and the
subjects are not related.

TYPES OF MOLECULAR PROBES:

MARKING PROBES:

Labeled probes (radiolabeled or immunolabeled) are used to visualize

the target molecules. For use with double-stranded DNA, it must first
be denatured by heat or high alkalinity. This labeled single-stranded
probe binds to the target; and the binding site is determined because
the labeling method leaves a signal that can be detected by
photography. In modern methods, fluorescent labels are used to emit
a signal that is detected by a laser, such as those used in real-time
PCR.

RADIOACTIVE:

Probes can be radiolabeled using nucleotides with specific atoms of a

radioactive isotope (usually 32P), but tritium is also used.

32P is usually used, since it is an isotope of phosphorus which, having

15 protons and 17 electrons (two electrons more than its isolated
element) produces the phenomenon of fluorescence when it comes
into contact with gasses in the atmosphere such as oxygen. Tritium is
also used, although in less proportion, since being radioactive it is
easy to locate, but also its radiation is not too harmful, since it
presents beta radiation.
ANTIBODIES:

Although radioactive labeling is very sensitive, it carries the dangers

associated with the use of radioactivity. Other antibody-based labeling
methods have been developed to avoid this danger. Using a
nucleotide-labeled probe attached to the molecule, specific antibodies
that bind to the molecule are added to the solution; these antibodies
are bound to a luminescent or fluorescent compound that leaves a
photographic imprint.