Bio-Rad DCode™ Universal Mutation Detection System - Manual (Page 60)

1x TBE Running Buffer
Reagent
Amount
10x TBE buffer 700 ml
dH
2
O 6,300 ml
6.3 Gel Volumes
The table below provides the required volume for the gel size and spacer thickness.
Spacer Thickness 20 x 20 cm Gel
0.75 mm 30 ml
1.00 mm 40 ml
6.4 Sample Preparation
1. It is important to optimize the PCR reaction to minimize unwanted products which may
interfere with gel analysis. The PCR products should be evaluated for purity by agarose
gel electrophoresis before being loaded onto an SSCP gel.
2. 150–300 ng of amplified DNA (usually 5–10% of the total PCR volume) can be loaded per
well. Aliquot the proper amount of sample into separate tubes and add equal volume of
2x SSCP gel loading dye. For extra control, the undenatured samples can be run on the
gel.
3. Denature the samples at 95°C for 5 minutes and then place on ice.
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