User:Paulina Alatriste/Notebook/UNAM Genomics Mexico 2011/2011/05/27

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TAP media preparation

ABSTRACT

  • Beginning of the preparation of TAP media for Chlamydomonas reinhardtii. Making of Beijerincks and Trace Elements solutions.

Today Pablo and I started to prepare TAP medium. This medium is necessary to grow Chlamydomonas reinhardtii, a single celled green alga that has a native system of hydrogen production. We are going to use it to compare this wild type system with the system engineered by our team and whose goal is to operate in R. etli.


We prepare the following stock solutions:


  • 1. Beijerincks solution 20X
NH4Cl 8g
CaCl2·H2O 1g
MgS04·H2O 2g

water to 1 liter

  • 2.Trace elements solution
EDTA disodium salt 50g
ZnSO4·7H2O 22g
H3BO3 11.4g
FeSO4·7H2O 4.99g
MnCl2·H2O 5.06g
CoCl·6H2O 1.61g
CuSO4·5H2O 1.57g
(NH4)6Mo7O24·H2O 1.10g

Dissolve each element separately: -H3BO3 in 200 ml


-ZnSO4·7H2O in 200 ml


-EDTA in 250 ml


-The rest of the substances in 50 ml


Mix all solutions except EDTA. Bring to boil, then add EDTA solution. The mixture should turn green. When everything is dissolved, cool to 70 degrees C. Keeping temperature at 70 adjust pH in a range of 6.5-6.8 with a solution of 20% KOH. Bring the final solution to 1 liter total volume. Stopper the flask with a cotton plug. The solution should eventually turn purple and leave a rust-brown precipitate, which can be removed by filtering.

    • We leave the flask resting all the weekend to let the precipitate formed. On monday we will finish the preparation of the media.