Column kits

The amplified cDNA template can be purified via phenol-chloroform-isoamyl alcohol extraction or with commercially available column kit. However, it was found that this purification step is not necessary for the following procedures. 

Zirconium columns kits for preparing affinity columns have been recently released by ZirChrom. They contain an activated linker that can be reacted with the target compound in the prepacked column to prepare the affinity column in situ. Generally, when an affinity column is made, the column must be dedicated to only that one separation. If you have six different affinity separations to make, you must buy six columns. But with these zirconium column kits, the affinity head can be stripped off in the column, the column cleaned, the linker reactivated, and a new affinity column created with a new affinity target without unpacking and repacking the column. This should open create new interest in affinity separations. 

Commercially available linear polymers and CGE kits are available. The chemical composition of some of these polymers is proprietary but others are not. Companies marketing these linear polymers for use in CGE include Bio-Rad,84 Perkin Elmer/ABI (linear polyacrylamide),85 and Beckman Instruments (PEO).86 The Beckman SDS-Protein Gel Column Kit includes many of the supplies and reagents required for Protocol 9. 

Silica-membrane column kit (QIAGEN DNeasy Plant Mini Kit) Anion-exchange column kit (QIAGEN Genomic-Tip Kit)... 

Hydrolysis of the stationary phase or other kinds of column aging may even occur during methods development. A finished method that needs to be used for a long time or by other workers should therefore always be verified on a new column, preferentially prepared from the same batch of packing. Column kits for this purpose are available fi om some manufacturers. 

During selection of these colunms, it may happen that on some columns two components are separated, but other components that are separated on other columns suddenly coincide. In such cases, POPLC can be used [17,18]. The principle is shown in Figure 4.11. A set of segmented columns is contained in a column kit. This set represents the colunms of differing selectivity described earher. From an initial measurement on the individual columns, the retention time of each... 

Each column is shipped with a PSS column quality certificate, a column connector, and the test kit, which was used at the PSS quality inspection laboratory to test the column resolution. This allows the user to reproduce the column tests without problems. It should be noted, however, that column test results not only depend on the performance of the column alone, but also on the instrument employed for the test. Figure 9.8 shows an example of a PSS column quality certificate. 

FIGURE 9.29 Analysis of natural spider silk (spider web). Columns PSS PFG 100 + 1000. Eluent HFIP + 0.1 M KtFat. Temp 2S°C. Detection UV 2S4 nm, Rl. Calibration PSS PMMA ReadyCal kit. 

One solution is to replace the column, but a less expensive approach is to attempt to clean the column. Baking is one approach that removes some forms of contamination, but also shortens the column life because it removes some of the stationary phase. A solvent rinse is the most effective means of cleaning a bonded or cross-linked phase column. Solvent rinse kits are available with instructions from most column manufacturers. The procedure involves forcing solvents through the GC column, usually in the following order—water, methanol, methylene chloride, and hexane—using 10-15 psi back pressure. 

Molecular weight distributions were examined by high performance size-exclusion chromatography (HPSEC) as described [22] on two serial Shodex OHpak KB-803 and KB-805 columns (0.8 x 30 cm Showa Denko, Japan) with a OHpak KB-800P guard column (0.6 x 5 cm), equilibrated at 1 mL/min in 0.1 M UNO3. The system was calibrated with a pullulan calibration kit (Showa Denko). 

Figure 5. Analytical isoelectric focusing. Ultrathin layers (0.4 nun) of polyacrylamide with ampholytes pH 2-11 were used. Samples of 10 pg of protein in 10 pi of 1 % glycine were applied. A.- Silver staining. B.- Stain for activity on overlays containing pectin in tris/HCl buffer at pH 8.0 with CaClj M.- Broad pi Calibration Kit protein (Pharmacia), samples of 5 pg of protein were applied. 1.-Ammonium sulphate precipitated proteins from cultures on pectin. 2.- Fractions with PNL activity eluted from the Superdex 75HR1030 column. 3.- Purified PNL.

Methods for DNA extraction Silica-membrane column-t)rpe kit Qiagen DNeasy Plant Mini kit Anion-exchange column-type kit Qiagen Genomic-tip kit CTAB method... 

Figure 8.17 Conversion of a standard gas chromatograph to a dual capillary column multidimensional chromatograph using a commercially available conversion kit. (Reproduced with permission from Scientific Glass Engineering). ...

RNA is then purified using column purification (we use the Ambion MegaClear kit, following the manufacturer s instructions). 

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