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Gabe15 · 06-17-2018, 05:46 AM

It could be coming from anywhere really. It is not enough to autoclave your tools if they are improperly handled afterwards. In fact, autoclaving your tools is not necessary as it's much better to flame/heat sterilize them immediately before you. The parts of the tools you use to manipulate the explants are they only part that needs to be sterile. Store your tools in ethanol in the hood to have on hand, and then hold the tips in the flame of an alcohol lamp to burn off the remaining alcohol and heat sterilize the tools. If you drop it, or touch something that wasnt sterile, re-sterilize immediately before using again.

You must also be sure your hood is functioning properly, that the filters are still good, and that you don't have dust accumulation in the venting system.

You also do not mention what type of plants you are using, or how you are preparing the material. Improper handling of the explants in the initiation phase is an extremely common and easy way of introducing contamination. How are you prepping and cleaning your starting material, and what kind of material are you using?

06-17-2018, 05:46 AM	#4 (permalink)
Gabe15 Moderator Gabe15's Photo Gallery Location: Oahu, Hawaii Find me on the map! Zone: 12 Name: Gabe Join Date: Jul 2005 Posts: 3,892 BananaBucks : 13,339,781 Feedback: 5 / 100% Said "Thanks" 1 Times Was Thanked 8,237 Times in 2,200 Posts Said "Welcome to Bananas" 8 Times	Re: Contamination problem It could be coming from anywhere really. It is not enough to autoclave your tools if they are improperly handled afterwards. In fact, autoclaving your tools is not necessary as it's much better to flame/heat sterilize them immediately before you. The parts of the tools you use to manipulate the explants are they only part that needs to be sterile. Store your tools in ethanol in the hood to have on hand, and then hold the tips in the flame of an alcohol lamp to burn off the remaining alcohol and heat sterilize the tools. If you drop it, or touch something that wasnt sterile, re-sterilize immediately before using again. You must also be sure your hood is functioning properly, that the filters are still good, and that you don't have dust accumulation in the venting system. You also do not mention what type of plants you are using, or how you are preparing the material. Improper handling of the explants in the initiation phase is an extremely common and easy way of introducing contamination. How are you prepping and cleaning your starting material, and what kind of material are you using? __________________ Growing bananas in Colorado, Washington, Hawaii since 2004. Commercial banana farmer, 200+ varieties.