Probe labeling is defined as sequence use to search the mixture of nucleic acid for molecule containing complementary sequence.In molecular biology, hybridization is a phenomenon in which single-stranded deoxyribonucleic acid (DNA) or ribonucleic acid (RNA) molecules anneal to complementary DNA or RNA

1. AMBER HASSAN
PhD Scholar
Aznostics the diagnostic center

2. *PROBE: A probe is a piece of DNA of the
sequence of the interest
*PROBE LABELLING: a labeled and defined
sequence use to search the mixture of nucleic acid
for molecule containing complementary sequence
*Probe concentration:
The higher the probe concentration, the faster
the hybridization occurs. However, the higher the
probe concentration, the higher the blot background
(i.e. probe will stick everywhere).
Low probe concentrations and long
incubations (overnight) usually produce the best
results.

3. LABELING OF DNAAND RNA
PROBES
*RADIOACTIVE LABELING: DISPLAY OR
MAGNIFY THE SIGNALSBY RADIOACTIVITY.
*NON RADIOACTIVE LABELING: DISPLAY
OR MAGNIFY THE SIGNALS BY ANTIGEN
LABELING.
*END LABELING: PUT THE LABELES AT THE
END.
*UNIFORM LABELING:PUT THE LABELS
INTERNALLY.

4. Classic procedure:
Radioactive labelling
of complementary
RNA

5. The Probe:
Artificial RNA
Labelled with DIG
Produced in vitro

7. *HYBRIDIZATION:
*In molecular biology, hybridization is a phenomenon in which single-
stranded deoxyribonucleic acid (DNA) or ribonucleic acid (RNA)
molecules anneal to complementary DNA or RNA.
*double-stranded DNA sequence is generally stable under physiological
conditions, changing these conditions in the laboratory (generally by
raising the surrounding temperature) will cause the molecules to
separate into single strands.
*These strands are complementary to each other but may also be
complementary to other sequences present in their surroundings.
Lowering the surrounding temperature allows the single-stranded
molecules to anneal or “hybridize” to each other.
*DNA replication and transcription of DNA into RNA both rely upon
nucleotide hybridization, as do molecular biology techniques including
Southern blots and Northern blots,the polymerase chain reaction
(PCR), and most approaches to DNA sequencing.

8. *1.DNA-DNA HYBRIDIZATION:

9. *2.FLUORSCENT IN SITU
HYBRIDIZATION:

10. *RNA/DNA hybridization is influenced by…
*Probe concentration
*
Sequence (GC bonds more stable than AT)
*Stringency