The document discusses different blotting techniques used in plant biotechnology. It describes the procedures and steps involved in Southern blotting, which involves restriction enzyme digestion of DNA, gel electrophoresis, transfer to a membrane, hybridization with a radioactive probe, washing, and autoradiography. The document also mentions Northern blotting and Western blotting but does not describe the procedures. Applications of Southern blotting include mapping restriction sites, DNA fingerprinting, and detecting transferred genes in transgenic plants.

1. PRINCIPLES OF PLANT
BIOTECHNOLOGY
Blotting
techniques
Prepared by; Kumar Harsh
25-09-2017
1
k.harsh

2. 25-09-2017
2
k.harsh

3. A. Southern blotting
Procedures;
1. DNA digestion through restriction endonucleases
2. Gel electrophoresis or PAGE
3. Alkali treatment
4. Buffer treatment
5. Transfer of material on to the nitrocellulose filter
membrane
25-09-2017
3
k.harsh

4. 6. Hybridisation with single stranded radioactive
DNA(probe)
7. Washing
8. Autoradiography
25-09-2017
4
k.harsh

5. Southern blotting
procedure;
25-09-2017
5
k.harsh

6. B. Northern blotting;
C. Western blotting;
25-09-2017
6
k.harsh

7. 25-09-2017
7
k.harsh

8. Applications of southern blotting;
1. It can be used to map the restriction sites around
a single copy gene sequence in any genome .
2. DNA fingerprinting
3. Preparation of RFLP maps
4. Detection and identification of the transferred
genes in transgenics.
Applications of nothern
blotting;
25-09-2017
8
k.harsh

9. Applications of western blotting ;
25-09-2017
9
k.harsh

10. Thank
you!!!!
25-09-2017
10
k.harsh
References:
a).Agarwal, P.K. & Dadlani, M. (eds.) 1987.
Techniques in seed science & Technology. South
Asian Publishers, New Delhi.
b) Agarwal, R.L. 1980. Seed Technology. Oxford &
IBH Publishing Co., New Delhi, India.