NORTHERN BLOTTING

1. NORTHERN BLOTTING Presented By : *M. UZAIR NIAZ *WAQAR ALI

2. Contents • What is Blotting? • Northern Blotting • History and introduction • Principles of Northern Blotting • Procedure • Advantages and disadvantages • Question answers

3. Blotting • Blotting is the technique in which nucleic acids or proteins are immobilized onto a solid support generally nylon or nitrocellulose membranes. • Blotting of nucleic acid is the central technique for hybridization studies. • Nucleic acid labeling and hybridization on membranes have formed the basis for a range of experimental techniques involving understanding of gene expression, organization, etc.

4. NorthernBlotting • Developed by James Alwine, David Kemp and George Stark in 1977. • Similarity of name with Southern Blotting but it analyzes RNA • To study gene expression by detection of RNA during differentiation, diseased conditions

5. Principle of Northern Blotting • Electrophoresis – Seperates RNA on the basis of their Molecular weight and type in agarose gel which have EtBr ,an intercalating agent in it. • Capillary action – RNA bands move towards blotting paper by capillary movement and entrap in sheet and buffer moves ahead.

6. Procedure

8. Diagrammatic View

9. Advantages • Simple method • Highly specific • Quality and quantity of gene can be measured Disadvantages • Detect small number of genes at a time • Use of ethidium bromide and UV light needs special training • Detection with multiple probes is difficult

10. THANK YOU