Southern Blotting-Immuno Blotting Techniques-P`ceutical Biotechnology-Unit 4- B. Pharmacy 6 Sem-L.02
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3 سال پیش
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The process of transfer of the denatured fragments out of the gel and onto a carrier membrane makes it accessible for analysis using a probe or antibody.
Depending upon the substance to be separated, blotting techniques may be– Southern blot, Northern blot or Western blot which separates DNA, RNA and proteins respectively.
Southern Blot is the analytical technique used in molecular biology, immunogenetics and other molecular methods to detect or identify DNA of interest from a mixture of DNA sample or a specific base sequence within a strand of DNA.
Principle of Southern Blotting
It is based on the principle of transfer of separated DNA fragments to a carrier membrane (usually nitrocellulose) using gel electrophoresis and subsequent identification of specific DNA fragments by labelled probe hybridization. Hybridization is a technique in which a double stranded DNA molecule is formed in between a single stranded DNA probe and a target single stranded DNA.
Procedure of Southern blotting
1. Extract and purify DNA from cells
2. Restriction Digestion or DNA Fragmentation
3. Separation by Electrophoresis
4. Depurination
5. Denature DNA
6. Blotting
7. Baking and blocking
8. Hybridization
9. Washing of unbound probes
10. Autoradiograph
Application of Southern blotting
Southern blotting technique is used to detect DNA in given sample.
DNA finger printing is an example of southern blotting
Used for paternity testing, criminal identification, victim identification
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The process of transfer of the denatured fragments out of the gel and onto a carrier membrane makes it accessible for analysis using a probe or antibody.
Depending upon the substance to be separated, blotting techniques may be– Southern blot, Northern blot or Western blot which separates DNA, RNA and proteins respectively.
Southern Blot is the analytical technique used in molecular biology, immunogenetics and other molecular methods to detect or identify DNA of interest from a mixture of DNA sample or a specific base sequence within a strand of DNA.
Principle of Southern Blotting
It is based on the principle of transfer of separated DNA fragments to a carrier membrane (usually nitrocellulose) using gel electrophoresis and subsequent identification of specific DNA fragments by labelled probe hybridization. Hybridization is a technique in which a double stranded DNA molecule is formed in between a single stranded DNA probe and a target single stranded DNA.
Procedure of Southern blotting
1. Extract and purify DNA from cells
2. Restriction Digestion or DNA Fragmentation
3. Separation by Electrophoresis
4. Depurination
5. Denature DNA
6. Blotting
7. Baking and blocking
8. Hybridization
9. Washing of unbound probes
10. Autoradiograph
Application of Southern blotting
Southern blotting technique is used to detect DNA in given sample.
DNA finger printing is an example of southern blotting
Used for paternity testing, criminal identification, victim identification
---------------------------------------------------------------------------------------------------------------------------------------------
My Telegram account: https://t.me/joinchat/UrMLkw_VPi5RMbO...
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My Facebook Page: https://www.facebook.com/MS-Sci-Acade...
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Click here to download Notes:
"https://forms.gle/8V2rRX4iHzhtAjxEA"
3 سال پیش
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