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Team:UEA-JIC Norwich/algaeglassbeads
From 2011.igem.org
1. Transfer 50ml of algae culture into four 50ml screw top tubes.
2. Place all four tubes in the eppendorf centrifuge at 800rpm for 9 minutes at 20-22°C.
3. Immediately pour contents of tubes into glass beaker and keep tubes upside down so no unwanted suspension mixes with the pellet.
4. For all tubes resuspend cells in 750µl of TAP media.
5. Wash pre prepared beads in ethanol removing excess gently.
6. Wash beads approximately 2-3 times in autoclaved distilled water.
7. Place approximately half the beads in a 1.5ml eppendorf tube for the control.
8. Resuspend both tubes with 200µL of distilled water.
9.Place samples from all four vials into 1.5 eppendorf tubes.
10.Centrifuge for 2 minutes at ~3000rpm.
11.Remove all excess supernatant.
12. Add 500µl TAP and re-suspend the pellet.
13.Cut tip off a 1ml pipette tip and add the 500µl of the suspended pellet to the tubes containing the beads.
14. To only one tube add ~1µl of Plasmid.
15. Place that tube on the vortex for ~20 seconds at 1800rpm, do the same for the control.
16.Add another 500µl TAP to the tube.
17.Perform a dilution of 1 fold and 10 fold for both the control and plasmid present tube.
18. Place 1ml of original sample onto TAP media plates.
19.Spead gently using a 1ml pipette tip.
20.Leave to dry and the incubate at 25˚C for 7 days in constant light.
