Re Mol Bio: Blotting Techniques

southern blot

discovered by edwin southern

southern blot

Target: DNA sample

southern blot

Probe: Nucleic acid

southern blot

Stain: Ethidium bromide under UV light

northern blot

Stain: Acridine orange, ethidium bromide

northern blot

Modification of the Southern blot technique

HCl

Depurination reagent

NaOH

Denaturation reagent

northern blot

Target: RNA sample

northern blot

Probe: Nucleic acid

western blot

Also known as the immunoblotting

western blot

Principle: Separation of fragments

western blot

Target: Proteins

beta-mercaptoethanol

Denaturation reagent

western blot

Stain: Amido black, Coomasie, Ponceau red

western blot

Probes: Proteins

Dot/Slot blots

Uses: expression,mutation, and amplification/deletion analyses

Dot/Slot blots

Target nucleic acid is deposited directly on the membrane and performed on less complex samples such as PCR products or selected mRNA preparations

Slot blots

more accurate for the quantitation

Dot blots

are useful for multiple qualitative analyses

Macroarrays

reverse dot blots on nitrocellulose membrane

Microarrays

used for improved spotting technology