( master mix was prepared an...)New Temperature Cycler Features Interchangeable Blocks

master mix was prepared, and 50-l aliquots were added to six thin-walled tubes. The tubes were positioned in the four corner wells and two center wells of the 40-well paddle of the Hot Top Assembly. A second set of reactions was prepared and cycled using identical PCR parameters and similar well positions using the 96-well block format and hot top assembly. Although not optimal for this primer-template system, an annealing temperature of 60C was used to allow both specific and extraneous PCR product bands to be compared. In addition, small deviations from 60C have been shown to produce discernible differences in banding patterns.³ After cycling with both block formats, 15-l aliquots of each reaction were electrophoresed through VisiGel separation matrix, stained with ethidium bromide and visualized using the Eagle Eye II still video system. Comparable product specificity and yield were observed when using either the 40- or 96-well block format (figure 2). Well-to-well variability was also low for both block formats, indicating that uniform temperature was maintained across the blocks during the amplifications. Similar results were observed with a different primer-template system (data not shown), indicating that consistent PCR results are obtained regardless of the block format used during experiments.

figure 2

Comparing the Hot Top Assembly to the Standard Paddle

PCR amplifications were performed to compare the product yield and banding pattern generated using either the Hot Top Assembly or standard paddle. The glucocerebrosidase primer-template system was again used for
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