Abstract: A system (10) for reducing volume of material used in laboratory processes, including a low-volume micro-plate (70,80) having a number of wells (14) bound together with matrix material (16), where the wells include secondary wells (74, 84) having reduced volume, and volume limiting plugs (20). The volume limiting plugs (20) have a shank portion (30) and a tip portion (28), where the shank and tip portions (20,30) are configured to extends a substantial distance into a well (14) of the multi-well plate (70,80) to reduce the volume of the well. Also volume limiting plugs (20) for use with standard multi-well plates and low-volume micro-plates (70,80) for use with standard plugs.

Abstract: In one aspect, the present invention concerns an improved method for the preparation of cDNA libraries. Preferred embodiments of the invention include methods and systems capable of generating cDNA libraries enriched for high molecular weight cDNA inserts. The method generally entails the following steps: (1) size-based separation of a plurality of mRNA molecules by Ion-Pairing Reversed-Phase Chromatography (IP-RPC), preferably using HPLC under denaturing conditions; and (2) collection of a fraction of the mRNA molecules that is enriched for mRNA molecules of a desired size range (in a preferred embodiment of the invention, larger-sized mRNA molecules are collected, e.g., mRNA of length greater than 10 kb). The collected fraction of mRNA molecules can be reverse transcribed to form a library of cDNA inserts enriched for inserts of a desired relative size range.

Abstract: The instant invention provides a non-HPLC chromatographic method for purifying a target polynucleotide comprising the steps of: applying the target polynucleotide to a separation medium having a non-polar separation surface in the presence of a counterion agent, whereby the polynucleotide is bound to the separation medium; eluting the target polynucleotide from the separation medium by passing through the separation medium an elution solution containing a concentration of organic solvent sufficient to elute the target polynucleotide from the separation medium; and collecting the eluted target polynucleotide. The separation medium can be supported in any of a variety of containers, non-limiting preferred examples of which include spin columns and vacuum trays. The invention is particularly useful for the separation of RNA and single and double stranded DNA.

Abstract: In one aspect, the present invention concerns an improved method for the preparation of cDNA libraries. Preferred embodiments of the invention include methods and systems capable of generating cDNA libraries enriched for high molecular weight cDNA inserts. The method generally entails the following steps: (1) size-based separation of a plurality of mRNA molecules by Ion-Pairing Reversed-Phase Chromatography (IP-RPC), preferably using HPLC under denaturing conditions; and (2) collection of a fraction of the mRNA molecules that is enriched for mRNA molecules of a desired size range (in a preferred embodiment of the invention, larger-sized mRNA molecules are collected, e.g., mRNA of length greater than 10 kb). The collected fraction of mRNA molecules can be reverse transcribed to form a library of cDNA inserts enriched for inserts of a desired relative size range.

Abstract: The instant invention provides a method for stabilizing an RNA molecule against degradation comprising applying a solution to a separation medium having a non-polar separation surface in the presence of a counterion agent, wherein the solution comprises the RNA molecule and an agent capable of catalyzing the degradation of RNA; eluting the RNA molecule from the separation medium by passing through the separation medium a mobile phase containing a concentration of organic solvent sufficient to elute the RNA molecule from the separation medium, where the elution is conducted under conditions that result in a substantial separation of the RNA molecule from the agent capable of catalyzing the degradation of RNA; and collecting an eluant fraction containing the RNA molecule that is substantially free of the agent capable of catalyzing the degradation of RNA. In a preferred embodiment the method is performed under conditions that are substantially free of multivalent cations.