Zone Electrophoresis (Brenner)

Date:
1959-1960
Reference:
PP/CRI/H/2/37
Catalogue details

Licence: In copyright

Credit: Zone Electrophoresis (Brenner). Source: Wellcome Collection.

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    M arme Biological LaLoratory WOODS HOLE. MASSACHUSETTS October l+ } I960 Dear Sydney, I have suceeded in separating host range mutants by zone electrophoresis. I thought you might be interested both in the mutants and in the technique. Included are several graphs. The mutants do not separate in the analytical ultra-centri fuge and the peaks of separate preparations are superimposable. Also,they do not separate in Tiselius electrophoresis. However, individual preparations give slight but significantly different mobilities. When the mobilities are translated into the geometry of the zone electrophoresis, they give the separation which is obtained. The latter is detectable only because of the sensitivity of the method. Run by gravity through a column of the same material of the same length over the same amount of time as used in the troughs, they show no separation. Furthermore, they seem to have different iso-precipitation points. The separation is,therefore, probably due to a charge difference. The mutants that have been tested are shown on the following page. The method was developed with Gy Levinthai and Simon Silver at M.I.T. The immobilizing agent is polyvinyl chloride powder M.Ik (Borden Mill^ Co. PVC 100, 40$/ lb.) which is made up like starch powder in 0.1 M acetate buffer pH 5»2. The runs are made at an applied volatge of 250 volts and 40 milliamps. The voltage across the troughs is usually 1Ö0 to 200 but this varies with the pre paration. The time is four hours. A: phage mixture of two or more