The results of an experiment comparing the sensitivity of electroblotting to vacuum blotting

The experiment was the following:
10 pg - 1 ng of a 6 kb plasmid was electrophoresed (0.5X TBE, 0.8% agarose) on two identical gels.

One gel was prepared for vacuum blotting (expose to UV, soak in 0.25N HCl, soak in 0.4N NaOH), and then transferred in a vacuum apparatus for 1 hour. After transfer the membrane was washed in 10X SSC, then UV-crosslinked and probed.

The second gel was transferred directly after electrophoresis by electroblotting for 1 hour at 12V in a Royal Genie.
After transfer, the membrane was washed in 10X SSC, then UV-crosslined, and then probed.

The same probe and washing conditions were used for both gels.


Self-evident. Electroblotting is not only faster, it is more sensitive.

Data generated by Chris Ott, Feinberg School of Medicine, Northwestern University.

Used by permission from Ashok Aiyar, Assistant Professor, Department of Microbiology-Immunology, Northwestern University