Is important for all Affinity Screening experiment templates. All wells that are part of a serial dilution should have the same Dilutionseries ID.Although TRIC is very robust against such buffer mismatches, it could affect the measurements. Take 1 mL of this dilution and add to the next tube (10 -2. Aseptically add 1 mL of enrichment sample to the first tube (10 -1) and mix gently. Label tubes 10 -1 to 10 -10 indicating dilution factor. Failure to do this, could bias the measured data with a gradient in salt, DMSO, glycerol or other additives. Using aseptic technique, dispense 9 mL of media into each of ten test tubes with sterile automatic dispenser or sterile 10 mL pipettes. The buffer in tube/well 1 and subsequent tubes/wells must be the same. Consider this step as the first dilution (10 1 ). Always mix small volumes with a pipette. Dilute the food sample 10× in diluent solution (e.g., saline solution or peptone water or peptone salt solution) ( see Note 1 ): 25 g (solid sample) or 25 mL (liquid sample) and add to the flask containing 225 mL of diluent solution ( see Note 4 ).Always use the smallest micro reaction tubes possible (e.g. The high surface to volume ratio leads to surface adsorption even for well-behaved proteins. The dilution factor (DF) of an X-fold dilution is X. 500 μl or more) and then transfer into the Dianthus microwell plate. For multiple serial dilutions, DF is equal to the product of dilution factors for each individual dilution. 20 μl) in large micro reaction tubes (e.g. Do not prepare less than 20 μl of sample.For ligands that are stored in DMSO, dilution is either performed in a way that maintains a constant level of DMSO throughout the dilution series or they are diluted in 100% DMSO and then each dilution point is diluted into buffer individually. The dilution factor can be freely determined by the user and is often adapted to obtain more data points for a transition phase or to cover a broader concentration range with fewer dilution points. This way, the ligand concentration is reduced by 50% in each dilution step. Typically, a serial 1:1 dilution (dilution factor 2 in DI.Control software) is performed by transferring one volume of ligand solution to an equal volume of buffer, mixing, and repeating this step.
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