Dear Mercy,
I would try to grow this cultures on MS0, if didn't do this so far. Very often it is the most simple and best option.
I am curious of results.....
wish you good luck
I recommend you use B5 medium (Gamborg); but not at the higher ionic strength (complete salts) use B5 medium to half its ionic strength (B5 / 2) added different concentrations of growth regulators and callus induction ,
I usually use MS/2 +0,5 mg/l 2,4-D + 1.5 mg/l Kent
Another important point is the concentration of the carbon source; we usually use concentrations from 3% to 0.5% sucrose
good luck
Dear Mercy
There are two phases for callus proliferation, once if you use MS salts complete strength you will have caulogenesis, and secondo for rhizogenesis, you use only half strength (MS/2 only for macroéléments). So, you have already callus formation, then you must not use 2.4D, but hormonal balance (auxine + cytokinine) because the role of 2.4D is callus induction which you have in you possession .
Good luck.
Dear Mercy,
I suggest, to try first increasing the concentrations of 2,4-D (0.5 or 1 mg/L) and adding a cytokinin, e.g, kinetin 0.1 or 0.5 mg/L. Good luck.
Hi Mercy,
Attached is a protocol for maintaining tobacco BY-2 cells. They use modified Linsmaier and Skoog (mLS) medium. See attachment.
MS Macronutrients, MS micronutrients (including iron..) + 200 mg/l KH2PO4 (up to 240 is OK) + 1 mg/l Thiamine HCl + 100 mg/l Myo-inositol + 0.2 - 1 mg/l 2,4-D (use less to avoid frequent subculture) + 30 g/l sucrose. pH 5.6-5.8. Incubation in dark . Make suspension also.
You can purchase media only with macro and macro : Sigma M5524, or phytotech
In the attached article, no need to add kanamycin. MSMO (MS with Minimal Organics)
Dear Mercy
I am a Ph.D. candidate at the University Of Mohaghegh Ardabili in Iran. My dissertation is to target plant organelles (mitochondrion and chloroplast) by functionalized carbon nanotubes.
I am looking for the seeds or cell lines of Nicotiana tabacum. L. cv Bright Yellow 2 [BY2].
In case you have access to this cultivar. I would like to kindly ask you to generously provide me seeds or cell lines of Nicotiana tabacum. L. cv Bright Yellow 2.
Dear Toraj Khabiri,
I have the cell line. I maintain it as callus. DMSZ germany also sells this cell line.
Best wishes
- Badges
- Science topic
- Similar topics
- Biological Science
- Biochemistry
- Biomolecules
- Peptides