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Tissue Culturing & Other Propagation Techniques of Banana Plants This forum is for discussing propagation techniques of banana plants. Tissue culturing is the popular process of creating clones from a source plant. There are other techniques to propagate banana plants however, such as nicking corms or dividing corms. Learn more inside.


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Old 06-12-2018, 04:23 AM   #1 (permalink)
 
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Default Contamination problem

I am trying to micropropagate, but there are usually contamination in jars.
I am performing intensive sterilization with NaClO and EyOH.
what else can I do?
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Old 06-12-2018, 11:21 AM   #2 (permalink)
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Default Re: Contamination problem

We need a lot more information on your technique before any useful advice can be given.

Do you use an autoclave or pressure cooker to sterilize your media? Do you have a functioning laminar flow hood or other sterile environment? Do you flame sterilize your tools before manipulation?
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Old 06-16-2018, 02:57 AM   #3 (permalink)
 
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Default Re: Contamination problem

Quote:
Originally Posted by Gabe15 View Post
We need a lot more information on your technique before any useful advice can be given.

Do you use an autoclave or pressure cooker to sterilize your media? Do you have a functioning laminar flow hood or other sterile environment? Do you flame sterilize your tools before manipulation?
I use an autoclave to sterilize the media and tools.
I transfer the plants to the media using laminar flow hood .
Within 1 week most plants die due to contamination.
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Old 06-17-2018, 05:46 AM   #4 (permalink)
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Default 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?
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Old 06-19-2018, 01:11 PM   #5 (permalink)
 
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Default Re: Contamination problem

Quote:
Originally Posted by Gabe15 View Post
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?

Thanks for your answer.
Our laminar cabinet is quite new.
Plant materials are G.nain. I use rhizomes as explant.
My surface sterilization work; 15% NaClO (30 min.) and 70% EyOH (5 min.).
The tools (penset etc.) are sterilized with Bead Sterilizer.
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