PCR-amplified DNA fragments were electrophoresed, prior to and following the cleanup process, on 1% agarose gels in Ix TBE buffer (50 mM Tris-HCl, 50 mM boric acid, and 1 mM EDTA) containing ethidium bromide, and visualized using a ChemiImager[R] 4400 Imaging System (Alpha Innotech, San Leandro, California).
If it is necessary to observe shorter fragments, a shorter running time would be needed as fragments less than 450 or so base pairs were run off the bottom of the gel when gels were electrophoresed for 11 hours.
T-cell receptor gene rearrangement analysis was performed by PCR in a multiplex reaction according to the method of Theodorou et al, (4) except that no GC clamps were added to the primers during their synthesis, and PCR products were electrophoresed on a 2.
They electrophoresed for one and one-half hours, stained the gel with ethidium bromide for 10 minutes and viewed and photographed the Hhal fragments with an ultraviolet light source for the gel image photo.
The protein lysate supernatants were electrophoresed on 15% (w/v) SDSpolyacrylamide gels, and unstained protein molecular weight marker (MBI Fermentas, Pittsburgh, PA, USA) was used as the molecular weight standard.