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=Amicon Ultra Centrifugal Filters= [[Daniel Jacobsen|Back to Main]] [https://www.emdmillipore.com/US/en/product/Amicon-Ultra-0.5%C2%A0mL-Centrifugal-Filters-for-DNA-and-Protein-Purification-and-Concentration,MM_NF-C82301?ReferrerURL=https%3A%2F%2Fwww.google.com%2F Amicon Order Page/Information] Amicon Ultra Centrifugal filters are microcentrifuge filters that allow you to purify and buffer exchange various MW cutoff biomacromolecules. They use a MW cutoff filter, say 10 kDa, to allow dialysate through while not passing the biomacromolecules. They can be used for buffer exchange via the following general protocol: <gallery perrow=2 heights=300px widths=400px> File:CentrifugalFilters-Purification.png|Purification/Concentration File:CentrifugalFilters.png|Buffer Exchange </gallery> ==Protocol-Filtration== <ol> <li>Add X uL sample and buffer to 500 uL total volume</li> <li>Spin down at 14000 x g for appropriate time (15 minutes for 22 uL volume)</li> <li>Discard flow through tube and get a new one</li> <li>Invert filter inside flow through tube and spin down at 14000xg for 1 minute</li> <li>Alternately, extract sample using pipette</li> </ol> ==Protocol-Dialysis== <ol> <li>Add X uL sample and new buffer to 500 uL total volume</li> <li>Spin down at 14000 x g for appropriate time (15 minutes for 22 uL volume)</li> <li>Discard flow through and add 500 uL new buffer</li> <li>Repeat steps 2 and 3 as many times as desired</li></ol> ===Dialysis Table=== {| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> |- style="background-color:#FFCC66;font-size:12pt;font-weight:bold" align="center" | width="120" height="30" | Starting Sample Volume | width="110" | 500 uL dilution | width="110" | Original Buffer Concentration | width="110" | Centrifuge Time |- style="font-size:12pt" align="center" | align="center" height="15" | 50 | align="center" | 10 | align="center" | 10% | 15 min |- style="background-color:#BFBFBF;font-size:12pt" align="center" | align="center" height="15" | 20 | align="center" | 25 | align="center" | 0.4% | 15 min |- style="font-size:12pt" align="center" | align="center" height="15" | 20 | align="center" | 25 | align="center" | 0.016% | align="center" | 15 min |} ==Spin Times and Concentration== [[Image:AmiconSpinTimeTable.png|800px]]
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