Hi everyone,
I am currently trying to get a root-organ culture going with field-collected spores. I am using a transformed chicorey root and MSR media. I am having problems with the surface sterilization of spores, probably due to insufficient duration, as i keep having bacterial growth in cultures. As a side thought, i started wondering, is there a good reason why i shouldn't add streptomcin sulphate to the MSR medium, to control any bacteria that might survive the main sterilization process? If this is ok, what concentration should i aim to use? I cant seem to find definitive literature on this.
Thank you in advance,
Tanel
Hi Tanel Vahter,
You will find your answer in this article.
Good Luck!
This type of bacteria growth medium is named by inventors deMann, Rogos, and Sharpe. Culture MacConkey designed to grow Gram-negative bacteria and differentiate them from antibiotics.
this one derives from Modified Strullu & Romand and is probably somewhat different. As I didn't find my answer from the article, I think I'm just gonna go for it and see what happens.
Dear Tanel,
We had same problem, but got some promising result by changing protocol for surface sterilization. Adding antibiotic is bitter experienced for us. we are surface sterling spores by using a solution (0.4% Cloramine-T +1-drops of Tween-20). Wash it with child water and and remove supernatant, repeat twice than re-suspend spores in 0.02% Streptomycine Sulphate for 10 min at fridge. Collect the spores after centrifucation and spread on M media . Incubate plates at 28 C temp and allow it to germinate. Used non contaminated and germinated spores for further application.
Note: Use laminar air flow and sterilized all reagent sterile with filtration and use chill.
- Badges
- Science topic
- Similar topics
- Agricultural Science
- Horticulture