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From 2012.igem.org

Contents 1 Mutagenesis 1.1 Washing column 1.2 Washing loop 1.3 Loading sample 1.4 Injecting sample onto the column 1.5 Washing column

Mutagenesis

Every buffer that you put into the FPLC must be filtered and be void of air to optimize the separation.

Washing column

1. Empty waste bin
2. Set loop valve on Load Position
3. Set column valve on designated position
4. Set on the pressure alarm: 5mPa
5. Set flow rate on half the maximum the column can handle
6. Run the FPLC for about 2 times the column volume
7. Empty waste bin

Washing loop

1. Set flow rate on 0
2. Set loop valve on Inject Position
3. Set column valve on Bypass position
4. Set on the pressure alarm: 5mPa
5. Set flow rate on half the maximum the column can handle
6. Run the FPLC for about 2 times the loop volume

Loading sample

1. Set flow rate on 0
2. Set loop valve on Load Position
3. Inject the sample into the loop with an syringe
4. Keep the syringe in the machine

Injecting sample onto the column

1. Set flow rate on 0
2. Set loop valve on Inject Position
3. Set column valve on designated position
4. Set on the pressure alarm: 5mPa
5. Set flow rate on half the maximum the column can handle
6. Run the FPLC for about 1 times the loop volume
7. Turn on the Frac 900, to collect the samples

Washing column

1. Set flow rate on 0
2. Empty waste bin
3. Set loop valve on Load Position
4. Set column valve on designated position
5. Set on the pressure alarm: 5mPa
6. Set flow rate on half the maximum the column can handle
7. Run the FPLC for about 2 times the column volume
8. Empty waste bin