Post Electrophoretic Analysis
Alkaline Blotting
One example of one might attempt alkaline blotting is listed here.
Alkaline Blotting: The Procedure
Positively charged nylon membranes allow the use of alkaline transfer buffers, which link the nucleic acids to the membrane without UV crosslinking. In some cases, alkaline blotting gives a higher background. Increasing the concentration of blocking reagent will often eliminate this problem.
NORTHERN BLOTTING
Follow the basic procedure given in the standard protocol for Northern Blotting, using 40mM NaOH in place of the 10X SSC.
SOUTHERN BLOTTING
Follow the procedure in the standard protocol for Southern Blotting, omitting Step 5 and using 0.4M NaOH in place of the 10X SSC.
NEXT TOPIC: Autoradiography
- UV Shadowing
- Uneven Staining
- Staining Proteins Immobilized on Membranes
- Staining Protein Gels with Coomassie Blue
- Southern Blotting
- Smeared Bands
- Silver Staining Protein Gels
- Silver Staining DNA Gels
- Protein Fixation on Gels
- Post-Electrophoretic Visualization with Nuclistain
- Overview of Western Blotting
- Northern Blotting
- Method for Western Blotting
- Mechanism of Immunostaining
- Mechanism of Immunostaining
- Immunostaining with Alkaline Phosphatase
- Guide Strip Technique
- Faint bands, low background
- Faint Bands, High Background
- Ethidium Bromide Staining
- Enzyme Linked Immunosorbent Assay (ELISA)
- Coomassie Blue Stain- Troubleshooting
- Blotches on Gel
- Autoradiography
- Autoradiographic Enhancement with Autofluor
- An Overview of Northern and Southern Blotting
- Alkaline Blotting