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There are two fundamental limitations on when it is possible to construct a lookup table for a required operation. One is the amount of memory that is available: one cannot construct a lookup table larger than the space available for the table, although it is possible to construct disk-based lookup tables at the expense of lookup time.
Proper column packing is important for maximum resolution: An over-packed column can collapse the pores in the beads, resulting in a loss of resolution. An under-packed column can reduce the relative surface area of the stationary phase accessible to smaller species, resulting in those species spending less time trapped in pores.
In contrast, most HPLC configurations use particulate packed columns; in these configurations, tiny beads of an inert substance, typically a modified silica, are used inside the column. [1] Monolithic columns can be broken down into two categories, silica-based and polymer-based monoliths.
Finally, elution is the process of adding an aqueous solution to the column, allowing the hydrophilic nucleic acid to leave the column and return to solution. This step may be improved with salt, pH, time, or heat. Finally, to capture the eluate/eluent, the column is transferred into a clean microtube prior to a last centrifugation step.