Recombination DNA Technology (Nucleic Acid Hybridization )

Nucleic Acid Hybridization

1. Nucleic acid hybridization YBT201 Submitted to presented by , Miss p. Mala M.Muthulakshmi Assistant professor 123011356013 Department of biotechnology M.Sc 1 year (Biotech) PMIST vallam

2. Contents Introduction DNA Hybridization probes Probe types Types of hybridization Application Reference

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. Types of hybridization There are mainly three techniques of hybridization 1. Southern hybridization 2. Northern hybridization 3. Colony hybridization

9. 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

10. 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

11. 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

12. Applications mRNA splicing studies To study the gene expression Of various tissue, organs , development stages, pathogen infection

13. 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

14. Applications Library screening Southern blot Northern blot ASOs to detect mutations

15. 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

16. 3) What is a probe A) Chemically synthesized DNA b) Purified DNA c) Fragmented DNA duplex d) Either purified or synthesized single stranded DNA Nucleic acid hybridization is process Used to identify Specific a) DNA b) RNA C) DNA sequence d) Nucleotides

17. 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

18. Answers C A D C d

19. Felsenfeld , G; Miles, HT (1967). “The physical and chemical properties of nucleic acids”. Apnnual Review of Biochemistry. 36: 407–48. McClean , Phillip. “Nucleic Acid Hybridizations”. DNA – Basics of Structure and Analysis. Retrieved 26 May 2017. Levsky , JM; Singer, RH (15 July 2003). “Fluorescence in situ hybridization: past, present and future”. Journal of Cell Science. 116 (Pt 14):

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