| Handling and Procedures
Before use, centrifugation of the deepwell Block with a swing-out rotor centrifuge is suggested, to remove solutions from the cover foil. Centrifugation prevents cross contamination while opening the plate. Otherwise drops under the cover foil may be shaken down. The film should be very gently peeled from the plate.
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First open the strap and turn it down to the surface of the deep well plate. |
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Then smoothly pull the strap in horizontal direction so that the aluminium foil moves parallel to the plate surface to avoid contamination of the wells by drops hanging on the foil. |
Instead of peeling the film, it can also be left intact and pierced to take out reagent.
The following procedure describes the use of this kit with the Sitting Drop Vapor Diffusion method.
For manual handling, either a 8 or 12 channel pipet can be used to pipet the recommended volume (50-100 µl) into the reservoirs of a 96 well crystallization plate. Pipet tips should be changed for each set of conditions. But several crystallization plates may be prepared at once (row or column wise) to save tips.
Transfer 50 nl to 2 µl of crystallization reagent from reservoir to the sitting drop well (for each well). Again a 8 or 12 channel pipet may be used. Pipet tips should be changed for each well or set of wells.
Pipet same volume of sample (50 nl to 2 µl) into each sitting drop well. Mixing can be performed by gently aspirating and dispensing the drop several times.
Seal the crystallization plate as you usually do or according to the suppliers recommendations. Seal the deepwell block using foils or cover mats (for short term storage). You may use the enclosed aluminium foils. Simply pull the aluminium foil from the protective film and put the foil on the plate by pressing it onto the surface with evenly spread pressure. Take care that the foil is flush with one side of the plate and overhanging on the other side. To simplify the reopening of the kit put a piece of adhesive tape on the underside of the overhanging aluminium foil.
Please note: When kits are sealed manually we highly recommend to store the kits in upright position at 4°C in the dark.
Sample solubility is also temperature dependent. Allthough most crystallizations have been achieved at room temperature, in many cases different temperatures have led to success. Comparison of results of screening at two different temperatures (4°C and room temperature) helps determine the magnitude of temperature effects on sample solubility. Temperature may be an important parameter in the optimization procedure.
Due to the standard format of deepwell-blocks, they are suitable for any type of standard liquid handler (working with single tip or needle up to 96 parallel channels).
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