- Mon Mar 17, 2014 10:10 pm
#198082
Hello all
I got seeds from the FTS on Monday(3/10), and have since put them onto agar. I am quite particular about my experiments, so I have everything documented:
3.7g/L of Now Foods Agar
1/3 MS
.1% PPM
pH 5.51
NO sucrose
Seeds were sterilized in Distilled water with 3% PPM for 4hours, 45minutes in a microcentrifuge tube, and rotated on a centrifuge tube rotator the entire time.
Media was sterilized using a research-lab grade autoclave for 15minutes heating, 15 minutes cooling.
Everything was handled in a laminar flow hood that had UV light on for 2 hours prior to opening, as well as 30 minutes air cycling before use to establish environment. 0% contamination so far.
The seeds were in the media on 2 sterile petri dishes, that have been parafilmed shut and have been under a large 125w CFL 6500K light( not for the light, but as of now, for the heat, and it has remained a constant 85 degrees). The photoperiod now is 24 hours long, and I will drop it down later, to 16h.
When these seeds germinate properly, and I have 2 cotyledons, I will transfer them to media with sucrose, as there was no need for them to have sucrose in the first batch
As of yesterday, 60% of seeds have germinated, which has me surprised because, that is pretty damn quick compared to other threads I have been following.
Photos will be uploaded tomorrow, when I have a chance, and as soon as I find a way to photograph with my dissection microscope.
I meant to start a thread a bit earlier than this, but got completely sidetracked.
I got seeds from the FTS on Monday(3/10), and have since put them onto agar. I am quite particular about my experiments, so I have everything documented:
3.7g/L of Now Foods Agar
1/3 MS
.1% PPM
pH 5.51
NO sucrose
Seeds were sterilized in Distilled water with 3% PPM for 4hours, 45minutes in a microcentrifuge tube, and rotated on a centrifuge tube rotator the entire time.
Media was sterilized using a research-lab grade autoclave for 15minutes heating, 15 minutes cooling.
Everything was handled in a laminar flow hood that had UV light on for 2 hours prior to opening, as well as 30 minutes air cycling before use to establish environment. 0% contamination so far.
The seeds were in the media on 2 sterile petri dishes, that have been parafilmed shut and have been under a large 125w CFL 6500K light( not for the light, but as of now, for the heat, and it has remained a constant 85 degrees). The photoperiod now is 24 hours long, and I will drop it down later, to 16h.
When these seeds germinate properly, and I have 2 cotyledons, I will transfer them to media with sucrose, as there was no need for them to have sucrose in the first batch
As of yesterday, 60% of seeds have germinated, which has me surprised because, that is pretty damn quick compared to other threads I have been following.
Photos will be uploaded tomorrow, when I have a chance, and as soon as I find a way to photograph with my dissection microscope.
I meant to start a thread a bit earlier than this, but got completely sidetracked.