CONTENT : Defination of blotting History Southern blotting Northern blotting Western blotting Application DEFINATION : Transfer of DNA or RNA or protiens fractinated by gel electrophoresis to a nitrocellulase paper or nylon mesh for separation , identification and analysis is called blotting or blot transfer . It is carried out by allowing a liquid to seep through electrophoresed gel , nitrocellulose paper and a pile of filter papers. They are very important for hybridisation experiments to determine the DNA , mRNA or protiens in electrophoresed gels , HISTORY In general , nylon membrane or nitrocellulose membrane is used for blotting tests . This technique was first devised ...
What is blotting? Blots are techniques for transferring DNA , RNA and proteins onto a carrier so they can be separated, and often follows the use of a gel electrophoresis. The Southern blot is used for transferring DNA, the Northern blot for RNA and the western blot for PROTEIN. TYPES OF BLOTTING TECHNIQUES Blotting technique Southern Blot Northern Blot Western blot It is used to detect DNAI.t is used to detect RNAIt. is used to detect protein. SOUTHERN BLOTTING • The technique was developed by E.M. Southern in 1975. • The Southern blot is used to detect the presence ...
2-D • Northern blot for RNA • Reverse Northern blot for RNA • Western blot for proteins • Far-Western blotfor Protein-Protein • Eastern blotting for posttranslational modification • Far-Eastern blot for Lipids, Drugs and Hormones • Dot blot • Slot blot ...
Blotting • All techniques use electrophoresis to separate. • All are based on hybridization technique • Difference in techniques lies in the target • Four applications –Western –Southern –Northern –Southwestern 08/27/2022 2 Hybridization The binding between ss labeled probe to a complementary nucleotide sequence on the target DNA. Degree of hybridization depends on method of probe labeling (radioacitve or non-radioactive system e.g. biotin-avidin. 08/27/2022 3 Probes • Labeled material to detect a target. • For DNA: 20-30 nucleotides, complementary to a region in the gene • Methods of labeling: • 32 Radioactive e.g. P ...
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