Re: [DIYbio] Tillandsia usneoides tissue culture and transforamtion

Nico, 

Im thrilled you want to delve into the dark-green arts! I have no experience with moss cept for physcomitrella patens but either 1-2mg/L of 2,4-D or a cocktail of 1mg/L BAP (6-benzylaminopurine) and 0.1mg/L NAA (napthaleneacetic acid) on MS media with gamborg b5 vitamins or with niacin at pH 5.8 is my go to for shot-in-the-dark tissue culture protocols. Replate every two weeks and until callus and then shift to just cytokinin and some giberellic acid to stretch out any stubborn shoots-to-be. Just go ease on the gibrellin since it can induce floral maturation in-vitro and force life cycle changes. Agrobacterium is my goto transformant but now with my shiny new gene gun ill be moving away from my little oncogenic friend and onto the tungsten pastures of biolistics. You can always harass Stan Gelvin at Purdue for his GV3101 agrobacterium tumefaciens strain (LHA4401 is decent but i prefer the more aggressive albeit harder to get rid of GV3101) or source it from your local university plant lab. Someone has to have it if they work with plants and genetics. Best of luck and dont give up!

Sebastian S. Cocioba

On Nov 11, 2015, at 3:41 PM, Nico B. <perpetuatetheimaginary@gmail.com> wrote: