3. INTRODUCTION
• It is the process in which two complementary single-stranded
DNA and/or RNA molecules bond together to form a double-
stranded molecule.
• Hybridization methods are based on labeled DNA or RNA acts
as probes to detect nucleic acids in specimens.
4. DNA
• Watson and crick model of DNA was followed And it possesses
Two strands
• Those strands are held by hydrogen bonds
• Nucleic acid hybridization is a process Used to identify Specific
DNA sequence . This is performed by means of DNA probe .
• Probes used In hybridization reactions are usually chemically
synthesised DNA or RNA
5. • Nucleic acid hybridization Can be done in all Combinations :
DNA-DNA, DNA-RNA Or RNA – RNA , Insitu hybridization And
FISH analysis
6. HYBRIDIZATION PROBES
• It is a nucleic acid Fragment that Is complementary To another
nucleic acids sequence
• A probe actually hybridize To single stranded nucleic acid
• Nucleic acid probe can be synthesised in the laboratory
7. PROBE TYPES
• Probes are three types
1. DNA probe
2. RNA probe
3. Oligonucleotide probe
8.
9. TYPES OF HYBRIDIZATION
• There are mainly three techniques of hybridization
1. Southern hybridization
2. Northern hybridization
3. Colony hybridization
10. SOUTHERN HYBRIDIZATION
• Southern blotting is a technique Employed for detection Of
specific DNA sequence in DNA sample
• It was First given by E.M Southern , in British biologist
• This method is include Separation of restricted DNA fragments
By Electrophoresis
11. APPLICATIONS
• Southern blotting used for Gene discovery, mapping, evolution
• It is used make RFLP maps
• Identify Specific DNA in the sample
• Used in phylogenetic analysis
12. NORTHERN HYBRIDIZATION
• Northern blotting was developed by James alwine, George
stark and Dravid kemp in 1977
• In this technique which RNA fragments Separated by
electrophoresis and immobilised on the membrane
• The Identification of specific RNA is done By using nucleic acid
probes
13. APPLICATIONS
• mRNA splicing studies
• To study the gene expression Of various tissue, organs ,
development stages,
pathogen infection
14. COLONY HYBRIDIZATION
• It is a method of Isolating a colony containing plasmid
Harboring a particular sequence Or gene from a mixed
population
• The colonies to be screened are first replic Plated onto a
nitrocellulose
16. 1) The process by which a probe is used to screen a library is
known as ___________
a) Hybridization
b) Southern blotting
c) Colony hybridization
d) Western blotting
2) In how many ways a DNA can be labeled?
a) 2
b) 3
c) 4
d) 5
17. 3) What is a probe
A) Chemically synthesized DNA
b) Purified DNA
c) Fragmented DNA duplex
d) Either purified or synthesized single stranded
DNA
4) Nucleic acid hybridization is process Used to
identify Specific
a) DNA
b) RNA
C) DNA sequence
d) Nucleotides
18. 5)Which of the following is not termed as hybridization?
A) DNA and cDNA
b) DNA and mRNA
c) DNA from different species
d) DNA from male and female of same species
20. 1. Felsenfeld, G; Miles, HT (1967). “The physical and chemical
properties of nucleic acids”. Apnnual Review of Biochemistry. 36:
407–48.
2. McClean, Phillip. “Nucleic Acid Hybridizations”. DNA – Basics of
Structure and Analysis. Retrieved 26 May 2017.
3. Levsky, JM; Singer, RH (15 July 2003). “Fluorescence in situ
hybridization: past, present and future”. Journal of Cell Science. 116
(Pt 14):
