I use phenol, SDS method to isolate RNA from plant cells, but I am confusing that how I can prepare sol, like TE, water satuarted phenol, which are free RNase. Just autoclaving after make solution ? I am going to use large amount so the glass bottle shoul dbe used.I am very afraid about RNA degradation that may cause by the botlles and water.Please give me some suggestions!
go for DEPC treatment... dip your glasswares, plastic ware etc. in 0.1% DEPC in molecular grade water ... all the best
1. Yes, as Prakash suggested, to prepare your experimental glassware with DEPC solution.
2. Prepare autoclaved DEPC-treated water, and use it to prepare other solutions.
3. Some people use RNase Decontamination Solution such as RNaseZap® to remove RNase from the working area, working tools (such as pipets) and equipment. I never used them. My RNA experiments still worked fine.
4. 'RNA degradation' sounds 'scary' to most beginners who start to deal with RNA. However, if you just follow the suggestions from an established protocol, you will be fine.
By the way, if you are interested, you can read the post from the link below:
"DEPC: The Wicked Witch of RNA?"
http://bitesizebio.com/9429/depc-the-wicked-witch-of-rna/
There is also a very good article regarding DEPC and RNA from ThermoFisher Scientific company. I recommend you to read it. The title of the article is:
"RNase and DEPC Treatment: Fact or Laboratory Myth"
https://www.thermofisher.com/us/en/home/references/ambion-tech-support/nuclease-enzymes/tech-notes/rnase-and-depc-treatment.html
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