Jump to content

  • Log in with Facebook Log in with Twitter Log in with Windows Live Log In with Google      Sign In   
  • Create Account

Submit your paper to J Biol Methods today!

(View All Videos) Toggle BioVideo

- - - - -

Handling RNA - How to avoid RNase contamination?

Hi Susan,

I would second what tea-test has pointed out. Any tissue you process is going to be loaded with RNases (cells contain RNases). Any residual RNase present on your homogenizer is going to be very minor compared to what is already present in your cells. All you need to do is wash your homogenizer with soap and water and give it a rinse with clean MilliQ or DEPC water.

In my experience working with RNA, the majority of the degradation happens at the initial stages of the isolation. You need to work quickly to get the cells lysed and homogenized within whatever lysis buffer you are using. Sometimes RNA degradation is a problem for very inexperienced users who are very slopping (touching the insides of tubes/lids with fingers even with gloves on, leaving lids open so that bacteria (that contains RNases) can float in, taking way too much of the top layer and getting the white goop (protein) when using Trizol etc.). That not withstanding, your biggest challenge will be to get the tissue homogenized quickly.

A note on RNases: Autoclaving is not effective at eliminating RNases. Check:

Also, even if autoclaving destroyed RNases, it would not be necessary to do. As tea-test pointed out, there should be very little RNases on your homogenizer, compared to what is present in your cells so autoclaving is not helpful here. Don't worry about using RNase Zap or any of that stuff on your homogenizer.

Another tip: When I do RNA work, I make sure I start with sterile/RNase free tubes. I take a whole bag and dump it out in the flow hood (not fume hood). Put on a clean pair of gloves. Cap all the tubes and store them capped in a jar. Now you have a clean supply of tubes ready to be used for any application, including RNA work. If you don't do this you may find yourself touching the inside of the caps when you close them when you go to label them. Your gloves will pick up RNases from the environment so don't consider them RNase free when you touch things.


I extract RNA from patients samples for detection of human viruses. We kept the samples in lysis-buffer (Qiagen RNA extraction kit) then later carry out extraction. Do you think it is a bad practice? Should we work immediately after adding samples to lysis buffer?

What is the sample? It all spends on if the lysis buffer is getting into the tissue/cells to protect the RNA from degradation. If it is tissue biopsy, the inner tissue that is not in contact with lysis buffer will have RNA degradation, that is why it is recommended to mince with scissors to less than 0.5mm thickness so that the sample can not only be coated in buffer, but it should be able to get to the inside portion of your sample.

I love this kind of republic audience and member of this forum who are so fearless and have a decent way to talk and help everybody like yesterday I was helped by a person who shared me https://www.proessay...om/essay-paper/ this awesome writing website to do all the things online without much into though

I remember this dissertation writing company was writing about this research. It was cool to read about it once again. I really enjoy reading some scientific content.

cHey! There you are! You can hire a subject professional for any type of work. Our grade miners writers specialize across all subjects and academic levels. Their profound expertise is the key to brilliant results and fast delivery. Any task is do-able with their pro help.

Thesis is a statement that lets the reader know what your position on the issue you are writing about. It must be precise and to the point. You should avoid any ambiguity in your statement as it might confuse the reader. An example of a correct thesis statement  you can find at https://freeessaywriters.net/ and it would be "the characters in" Kill a Mockingbird "suffer great injustice because of racism and prejudice."

Home - About - Terms of Service - Privacy - Contact Us

©1999-2013 Protocol Online, All rights reserved.