Team:Wageningen UR/Protocol/FPLC
From 2012.igem.org
(Difference between revisions)
MarkvanVeen (Talk | contribs) (→FPLC user manual) |
MarkvanVeen (Talk | contribs) (→FPLC user manual) |
||
Line 4: | Line 4: | ||
'''Every buffer that you put into the FPLC must be filtered and be void of air to optimize the separation.''' | '''Every buffer that you put into the FPLC must be filtered and be void of air to optimize the separation.''' | ||
- | == | + | == Washing column == |
<ol> | <ol> | ||
<li>Empty waste bin</li> | <li>Empty waste bin</li> | ||
Line 15: | Line 15: | ||
</ol> | </ol> | ||
- | == | + | == Washing loop == |
<ol> | <ol> | ||
<li>Set flow rate on 0</li> | <li>Set flow rate on 0</li> | ||
Line 25: | Line 25: | ||
</ol> | </ol> | ||
- | == | + | == Loading sample == |
<ol> | <ol> | ||
<li>Set flow rate on 0</li> | <li>Set flow rate on 0</li> | ||
Line 33: | Line 33: | ||
</ol> | </ol> | ||
- | == | + | == Injecting sample onto the column == |
<ol> | <ol> | ||
<li>Set flow rate on 0</li> | <li>Set flow rate on 0</li> | ||
Line 44: | Line 44: | ||
</ol> | </ol> | ||
- | == | + | == Washing column == |
<ol> | <ol> | ||
<li>Set flow rate on 0</li> | <li>Set flow rate on 0</li> |
Latest revision as of 07:40, 11 September 2012
Contents |
FPLC user manual
Every buffer that you put into the FPLC must be filtered and be void of air to optimize the separation.
Washing column
- Empty waste bin
- Set loop valve on Load Position
- Set column valve on designated position
- Set on the pressure alarm: 5mPa
- Set flow rate on half the maximum the column can handle
- Run the FPLC for about 2 times the column volume
- Empty waste bin
Washing loop
- Set flow rate on 0
- Set loop valve on Inject Position
- Set column valve on Bypass position
- Set on the pressure alarm: 5mPa
- Set flow rate on half the maximum the column can handle
- Run the FPLC for about 2 times the loop volume
Loading sample
- Set flow rate on 0
- Set loop valve on Load Position
- Inject the sample into the loop with an syringe
- Keep the syringe in the machine
Injecting sample onto the column
- Set flow rate on 0
- Set loop valve on Inject Position
- Set column valve on designated position
- Set on the pressure alarm: 5mPa
- Set flow rate on half the maximum the column can handle
- Run the FPLC for about 1 times the loop volume
- Turn on the Frac 900, to collect the samples
Washing column
- Set flow rate on 0
- Empty waste bin
- Set loop valve on Load Position
- Set column valve on designated position
- Set on the pressure alarm: 5mPa
- Set flow rate on half the maximum the column can handle
- Run the FPLC for about 2 times the column volume
- Empty waste bin