Re: TTC test


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Abgeschickt von Michael Weinert am 05 November, 2003 um 23:13:19

Antwort auf: Re: TTC test von Bill Steele am 05 November, 2003 um 23:10:58:

Dear Bill,

The TTC test is carried out as follows:

After the bleaching procedure the seed has to be thoroughly rinsed with sterile water (otherwise false positive results possible by staining microorganisms). Then the seed is immersed in 10 ml TTC solution. Important: All glass vessels containing TTC solution have to be kept dark by wrapping them with aluminium foil, otherwise the colour reaction starts. Incubate the TTC test 5 days at 25°C (77°F), 1-2 days as normally recommended are too short. After this rinse the seed with tap water to avoid any further coloration by the effect of light and place the seed on a filter paper for counting. Once the colour reaction has taken place, the coloration is not sensitive to light.

Making of the TTC solution:

dissolve 0.36 g KH2PO4 in 50 ml aqua dest.
dissolve 1.43 g Na2HPO4 x 12 H2O in 50 ml aqua dest.
mix these two solutions and add 1.0 g 2,3,5-Triphenyltetrazolium chloride (keep the solution dark in the fridge as it is sensitive to light)

I would not recommend to use old TTC solutions or old TTC salt. The reaction is pretty sensitive and old TTC has lost its ability to react.


I hope this helps

Michael


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