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separation Buffers are often cooled and stirred to minimize i2 R heating and local pH changes that may occur near the gel buffer interface When the tracking dye has migrated to the bottom of the column, the gel is removed using pressure, and proteins are xed in place by precipitation [this occurs during a soaking step in a solution of trichloroacetic acid (TCA)] and are then stained for detection Slab PAGE allows simultaneous electrophoresis of a number of samples to be performed under identical conditions Slab PAGE has a higher resolving power than column PAGE, and is often used for molecular weight and purity determinations Because the slab is relatively thin, heat dissipation is more ef cient in a slab than in a column.

16

When disks are shared, SAN-style, between the nodes in an SDC, the data need only be written once, twice when mirroring is in place, with all writes performed locally (Disk mirroring does add a small amount of performance overhead, but in most cases, that overhead is not noticeable and is certainly not significant) But the real problem with RDCs is what happens after a failover Consider two hosts, eatdrink and bemerry, configured asymmetrically (only because it s simpler; the principles would apply equally in a symmetric or a larger SANbased cluster configuration) in an RDC that uses software replication Eatdrink, the primary, is working and servicing its users, and replicating data over to bemerry, the takeover host When eatdrink fails, bemerry takes over; its data is an up-to-date and current copy of what was on eatdrink before it failed.

The slab can be used vertically or horizontally, but in practice, the horizontal slab method is used only when the monomer plus cross-linker concentrations are low and the gel is soft A slab gel is prepared in the same way as a column gel, except that a comb, or slot-former, is inserted before polymerization Removal of the comb after the gel has set leaves sample wells that are separated from each other by continuous strips of gel In zone electrophoresis, two components are considered separated when the difference in migration distances exceeds the width of the original sample band Because of diffusional band broadening, the sharpness of bands diminishes with increasing run time.

During the time that eatdrink is down, writes that reach bemerry do not get replicated back to eatdrink because there is no way for eatdrink to process incoming data packets When eatdrink is fixed, it cannot take over for bemerry until its disks have been refreshed with the changes that occurred on bemerry while it was down This refresh can require manual intervention and will certainly require time and network bandwidth before it is completed Let s also consider what would happen if during the time when eatdrink is being refreshed with the changes from bemerry (or before that refresh has begun), there is, for example, a power failure that shuts down both systems Bemerry is the only system with current data; the data on eatdrink is still out-ofdate It is difficult and may well be impossible for clustering software to determine which host s data is more current.

A method used to improve the resolving power of zone electrophoresis that is unique to gel media involves a stacking gel between the sample and the running gel, and is called disc electrophoresis,5 due to the electric eld strength discontinuity that yields a very narrow sample zone Figure 99 shows the components of the gels and buffers used for this method The running gel in disc electrophoresis is cast in the presence of the running buffer, typically a high pH glycine buffer The stacking gel is cast in the presence of a tris buffer at lower pH and ionic strength Because the monomer concentration is much lower in the stacking gel, frictional drag is lower and analyte mobility is higher The stacking gel improves resolution because of the pH discontinuity, and the buffering effect that occurs at the interface between the running and stacking gels At pH 6.