View Single Post
Old 09-01-2012, 11:24 AM   #9 (permalink)
jmoore
 
jmoore's Avatar
 
Location: Lowestoft, UK
Zone: 8a
Name: James
Join Date: Aug 2008
Posts: 766
BananaBucks : 118,593
Feedback: 4 / 100%
Said "Thanks" 192 Times
Was Thanked 799 Times in 373 Posts
Said "Welcome to Bananas" 71 Times
Default Re: !st time Tissue Culturing with pictures

Your previous linked post shows you are using the correct part of the plant.

As for the contamination. Could be anything.

Do you add a surfactant to your bleach solution, like tween 20? it reduces the surface tension and allows deeper penetration.
Do you rinse 3 times in sterilsed water?
I used a dip in 70% Iso Propyl alcohol before cutting the meristem in the hood.
Is the hood sterilised? Spraying with IPA before use is a good idea.
Do you sterilise your tools before and after each use? They should be run through a spirit lamp or dipped in 70 % IPA before and after each use.
Did you wash your hands and forearms?
Your arms should be bare so as not to introduce anything from your clothes.
You should disinfect your hands with an alcohol based hand cleaner or wear sterilegloves (whichever is the cheaper)
Do you add PPM to your medium? Plant preservation material is a broad spectrum microbicide and will help combat mild contamination. It should be added at 1 mg/l.
Do you use sterile paper towels. Cutting should be done on sterile paper towels.

I said it could be anything.
jmoore is offline   Reply With Quote Send A Private Message To jmoore
Said thanks: