OGMP - Sequencing Gel Electrophoresis

(Version 21-MAY-1995)
We have developed a high resolution gel system which is fast and easy to handle, reliable and inexpensive. The principles are the following:

Very thin (0.2mm), 60 cm long acrylamide gels; fast plate assembly due to spacers permanently fixed to one plate; easy pouring of gels without sealing of glassplates; reduction of band distortion (smiling) through an aluminium plate that distributes the heat in the gel sandwich; simple, self-made chambers; equalization of band spacing using an external buffer gradient; no manoeuvring of gels prior to exposure, instead, in situ dry-fixation of the gel matrix onto the glassplate without chemical binding.

The method has been published (BF Lang & G Burger 1990 Anal Biochem 188:176-180) and a few modifications have been made since. The most important are that we now use mostly 4% (sometimes 5%) acrylamide, 1/8xTBE-buffer in the upper and 1xTBE + 0.1 M potassium phosphate in the lower tray.

We read from one film on average 500 nts (counting from the SmaI cloning site of pBluescript), nt.1-350 with 100% accuracy and the following 100 - 200 nts with 98% accuracy.

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