In general, autoclaving will NOT deactivate RNases. More importantly, you should also note that although you can use DEPC to treat glassware, DEPC is usually used to treat water or aqueous solutions, NOT for glassware. Glassware should be baked at 180C for 8 hours. As for plastic-ware, it is usually easier to simply buy RNase-free disposable plastic-ware. If there are large pieces of non-disposable plastics that you NEED to use, I suggests something like this:
http://www.lifetechnologies.com/order/catalog/product/AM9786
I emphasize NEED. It is often easier to simply find a protocol that lets you use all disposable plastics. I think we could guide you more appropriately if you gave us a better idea of what you want to do. For the most part, RNA mini-preps for RT-PCR purposes are all done in disposable plastics. There is some good information here:
https://www.neb.com/tools-and-resources/usage-guidelines/avoiding-ribonuclease-contamination
In general, autoclaving will NOT deactivate RNases. More importantly, you should also note that although you can use DEPC to treat glassware, DEPC is usually used to treat water or aqueous solutions, NOT for glassware. Glassware should be baked at 180C for 8 hours. As for plastic-ware, it is usually easier to simply buy RNase-free disposable plastic-ware. If there are large pieces of non-disposable plastics that you NEED to use, I suggests something like this:
http://www.lifetechnologies.com/order/catalog/product/AM9786
I emphasize NEED. It is often easier to simply find a protocol that lets you use all disposable plastics. I think we could guide you more appropriately if you gave us a better idea of what you want to do. For the most part, RNA mini-preps for RT-PCR purposes are all done in disposable plastics. There is some good information here:
https://www.neb.com/tools-and-resources/usage-guidelines/avoiding-ribonuclease-contamination
i agree with Mr.Mark, rinse in chloroform also gives better results for plastic and glassware followed by autoclaving. as i usaully practice in RNA extraction protocol.
Yes definitely you have to wash or rinse every equipment with DEPC before going for RNA extraction. RNAse can degrade the RNA..You need to take lots of precaution before and during RNA extraction..
Well it also depends on the purpose of the extraction. But like everyone has mentioned it its very important to have RNAse free equipment for extraction and isolation. The best would be to use Nuclease free plasticware which is the the normal way of working.
@Sudheer: Be careful with Chloroform, there is plasticware around, which doesn't like to get in contact with it (it would be useless in RNA extraction, since the solutions contain chloroform).
Autoclaving will not destroy all RNase, though it may inactivate a portion of it. Its true that Autoclaving denatures RNase but many RNase are quite capable of renaturing back to active form once the solution cools down. It is always safe to use 0.1% DEPC treated utensils and solutions for RNA isolation.
@Christian Praetorius : but we need to autoclave after wash with chloroform sir as i use it regularly but i have to take care as u said thanks for the suggestion sir
Yes exactly require because DEPC water doesn't have RNases even though it present as inactive form.But one limitation is that after autoclave there is chances to have minor traces of DEPC present, which will interfere in your RNA complete extraction by forming caroxy methylation process.
There is a really cool material on life tecnologies site showing the difference of using or not DEPC on your water. Some of the ideas there are analog to plastic and glassware cleaning. Though I would sugest the proceadures Mark mentioned above.
http://www.lifetechnologies.com/br/en/home/references/ambion-tech-support/nuclease-enzymes/tech-notes/rnase-and-depc-treatment.html
I agree with mark Baking glass at 180°C for 5-8 hours is sufficient, plastics from a new bag is even better than autoclaving it.
If you prefer Hot phenol extraction method, there is no need of any baking or DEPC
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