Glass Bead Transformation of Chlamydomonas
Grow cell culture to an OD750 of 0.15 to 0.4 .
Centrifuge at 400g for 5 minutes at room temperature .
Resuspend in 1 / 100th the original volume of TAP .
Add the following : 8000 , PEG to 5 % final conc . , 3 ug of DNA , 0.3g of 500 , micron glass beads 0.4mL , Chlamy cell suspension .
Mix with a pipette Vortex at max speed for 15 seconds .
Take 25uL of the cell suspension and add to 100uL of TAP with an appropriate antibiotic .
Spread on a TAP or YA plate , with an appropriate antibiotic , using large glass beads .
Allow the liquid to dry while avoiding light .
Seal the plates with parafilm .
Allow the colonies to grow ( colonies will appear in 1 - 3 weeks ) .
Transfer the remaining cell / vortex culture to a 125mL flask with 20mL of TAP .
w / o antibiotic Incubate for 6 hours on an orbital shaker at 70rpm .
Add antibiotic to an appropriate concentration .
Take 50uL of the cell suspension and spread on a TAP or YA plate with an appropriate antibiotic with large glass beads
