Daniel:Protocols/AmiconUltra: Difference between revisions
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>Djacobse (Created page with "=Amicon Ultra Centrifugal Filters= Back to Main [https://www.emdmillipore.com/US/en/product/Amicon-Ultra-0.5%C2%A0mL-Centrifugal-Filters-for-DNA-and-Prot...") |
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[[Image:CentrifugalFilters.png|400px]] | [[Image:CentrifugalFilters.png|400px]] | ||
==Protocol== | ==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><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> |
Revision as of 21:43, 13 May 2017
Amicon Ultra Centrifugal Filters
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:
Protocol-Filtration
- Add X uL sample and buffer to 500 uL total volume
- Spin down at 14000 x g for appropriate time (15 minutes for 22 uL volume)
- Discard flow through tube and get a new one
- Invert filter inside flow through tube and spin down at 14000xg for 1 minute
- Alternately, extract sample using pipette
Protocol-Dialysis
- Add X uL sample and new buffer to 500 uL total volume
- Spin down at 14000 x g for appropriate time (15 minutes for 22 uL volume)
- Discard flow through and add 500 uL new buffer
- Repeat steps 2 and 3 as many times as desired