FlyTrapCare Forums

Hi rellenburg,

I'm sorry but my english is bad
I put into autoclave all tubes and caps but the medium was into a autoclavable bottle(2 x 500ml).

Thanks for the info..I really have to change some products concentration and the time to expose the tissue.

So I'll disolve my H2O2 to get 3%.
I'll write the news soon.

Best regards

Hey friends,

Following the sterilization methods:

I just bought a soap with triclosan but how can I do a solution with it?
Only dissolve it in sterilized water?

Many thanks

Ricardo

I use it as a first clean step, so I just use tap water. I fill a 4oz baby food jar with tap water and 1 drop of soap. Shake for 2 min. Pour explants into a tea ball. Rinse under running tap water for 3 min. Then procede to the chemicals. I'm sure others do it differently.

Since that soap normally has that hormonal disruptor in it, and it is in a bottle with a "applicator" I press it 2 times (about 3ml I think) for a 150ml jar with tap water, shake and rinse in almost sterile distilled water.

Haha. I knew someone would have a different method I've never used it that strong.

Hello,

Ok I'll mix 3ml of my liquid soap in 150ml distiled water hehehehe.

I want to try with Drosera capensis(8cm leaf), Drosera burkeana(2cm leaf) and vft wacky trap(1-2cm leaf)

How long time I have to expose this explants in yours opinion?

Rellenburg / Coline >>> Thanks a lot!

Regards
Ricardo

The soap is not very harmful since what I have seen, so any time between 5-30min

Ok I'll do!
Thanks

Let us know how it goes for you. I have not experimented with the length of time with the soap, so I really can't give any other advice than what I've posted already. The majority of my failures result in dead tissue, not contamination, so I would be reluctant to go longer in any step. Unless of course I went longer in the soap and backed off some time in the bleach step. Hmmmm, may ahve to try that. Good luck!

Can I use a magnetic stirrer to make the sterilization?
I can do it easily....but I don´t know if this method would be good, maybe reduce the time to expose....

Highlander wrote:Can I use a magnetic stirrer to make the sterilization?
I can do it easily....but I don´t know if this method would be good, maybe reduce the time to expose....

Yes, I successfully use a magnetic stirrer in the cleaning of explants or seed. Generally, I use the tiniest ones, about the size of a pencil's eraser, which will fit within small 50 ml beakers or even flat-bottomed test tubes . . .

I have baby food jars(30-40ml) and beckers(50ml)

I just read that my liquid soap has 1% triclosan so I have to add 1 or 2 drops into 50ml dest. water? is ok?

Thanks
Regards

Highlander wrote:I have baby food jars(30-40ml) and beckers(50ml)

I just read that my liquid soap has 1% triclosan so I have to add 1 or 2 drops into 50ml dest. water? is ok?

Thanks
Regards

Yes, that soap will work; but definitely wash the explant under running water first, for five minutes at the very least, before soaking . . .

I did a step by step to make my sterilization method as described bellow:

I read the FAQ and the answers on this topic, so I made (at home) 4 main chemical solutions to try here.

1) Triclosan solution( a bottle with 1 liter of distilled water + 1full cap of liquid soap 1% troclosan);
2)H2O2 3% solution ( 3 flasks with 50ml bought in a drugstore);
3)NAClO 10% solution + tween20 ( a bottle with 900ml of distilled water + 100ml of bleach + 40 drops of tween20);
4)PPM solution (a bottle with 1 liter >>> ppm 1ml/L);

I want to do this method with 4 or 5 leafs from my vft , each about 2-3cm lenght.
All steps will be made with a magnetic stirrer and jars about 40-50ml(except running water) to clean the explants.

1) wash 10 min. in running water
2) wash 10 min in triclosan solution
3) wash 5 min in distilled water
4) wash 2 min in H2O2 3% solution
5) wash 5 min in distilled water
6) wash 5 min in NAClO 10% + tween20 solution
7) wash 5 min in distilled water
8) wash 10 min in PPM solution
9) wash 5 min in distilled water
10) wash 5 min in distilled water

I'm waiting for suggestions because I think that will make this process until the next weekend.

Thanks in advance

You are killing any kind of effect that PPM gives to the culture using Tween, the manufacturer says that
For everything else it is a good procedure