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I use RNase AWAY in the lab. I would like to know how dangerous this chemical is for health. For example, when I remove my gloves my hands smell because of the RNAse AWAY
RNase AWAY is marked for Category II skin corrosion/irritation and Category II eye irritation/serious eye damage. Recommendations by the MSDS followed by definitions as follows:
Skin contact Wash off immediately with plenty of water for at least 15 minutes. Remove and wash contaminated clothing before re-use. Immediate medical attention is required.
Eye contact Rinse immediately with plenty of water, also under the eyelids, for at least 15 minutes. Immediate medical attention is required.
DEFINITIONS FOR GHS LABELING
More information is included in the MSDS provided.
As for any smell, RNase away is reported to be a proprietary alkali hydroxide solution, so without any additional information it's difficult to tell.
If you are doing RNA preps, there isn't much of a way around it. RNase are everywhere and the only way not the degrade your sample is to use proper technique, barrier protection such as nitrile gloves, certified RNase free containers (macrophage tubes, pipet tips, etc.), and saturate your work surface and instruments with RNAse away.
If you really have a concern and it is available to you, a BSL-2 tissue culture hood with laminar air flow, that you get permission to use for the purpose of an RNA prep, is an option for an added level of safety. You still have to prepare your work surface with it, but the laminar air flow will keep any fumes away from you. A regular fume hood would probably be enough of a precaution, again if you are concerned, and it is less likely that anyone would take issue to you working in it if the BSL-2 hood is set aside for the exclusive use of tissue culture work.
You should, in general read the Material Safety Data Sheet for whatever chemical you are using. This is always provided by the vendor.
For RNAse-AWAY (Sigma-Aldrich):
Gloves should be worn when handling this product. RNase Decontamination Reagent is alkaline in nature and will cause irritation if prolonged contact with the skin is allowed. In case of contact with eyes, immediately flush with water for fifteen minutes and contact a physician. If swallowed, do not induce vomiting. Give plenty of water and contact a physician immediately.
According to the MSDS:
Not a hazardous substance or mixture according to Regulation (EC) No. 1272/2008. This substance is not classified as dangerous according to Directive 67/548/EEC.
First Aid measures as per the MSDS.
If breathed in, move person into fresh air. If not breathing, give artificial respiration.
In case of skin contact
Wash off with soap and plenty of water.
In case of eye contact
Flush eyes with water as a precaution.
Never give anything by mouth to an unconscious person. Rinse mouth with water.
Coordinating the removal of RNA-DNA hybrids
Credit: University of Mainz
Two research teams led by Professors Brian Luke and Helle Ulrich at the Institute of Molecular Biology have deciphered how two enzymes, RNase H2 and RNase H1, are coordinated to remove RNA-DNA hybrid structures from chromosomes. RNA-DNA hybrids are important for promoting normal cell activities like gene regulation and DNA repair, but having too many is also a risk for DNA damage and can lead to neurodegenerative disease and cancer.
In their article, which was published today in Cell Reports, Luke and Ulrich show that the enzyme RNase H2 removes RNA-DNA hybrids primarily after DNA replication. Any remaining RNA-DNA structures are then removed by RNase H1, which acts independently of cell cycling.
DNA is normally found as a stable, double-stranded structure. However, DNA also sometimes interacts with RNA to form RNA-DNA hybrid structures that regulate gene expression and DNA repair. R-loops are a special type of RNA-DNA hybrid in which an RNA strand binds to one strand of a DNA molecule and pushes out the other DNA strand so that it is exposed as a single-stranded loop. R-loops can regulate gene activity, but also quickly become dangerous because incorrect removal can damage the DNA, potentially causing mutations. Therefore, excess R-loop formation can be toxic for cells—indeed, mutations in R-loop removal proteins are known to contribute to neuroinflammatory diseases and cancer.
R-loop removal is catalysed by the enzymes RNase H1 and RNase H2, which degrade the RNA strand. In addition, RNase H2 also has a secondary ability to excise single ribonucleotides, which can sometimes be mistakenly incorporated into DNA by polymerases in a process known as ribonucleotide excision repair (RER). Previous studies showed that mutation of RNase H2 disrupted genome stability more than RNase H1 mutation, suggesting that RNase H2 has a more important role in maintaining genome stability. However, it was never fully understood how these important enzymes are coordinated.
To dissect the distinct roles of RNase H1 and H2 in R-loop removal, the Luke lab engineered yeast to express RNase H1 and H2 only during specific phases of the cell cycle and then exposed them to methyl methanesulfonate (MMS), an agent that increases R-loop formation. Only yeast that could effectively remove R-loops would survive, while those with impaired R-loop removal would not survive.
With support from the Ulrich lab, they found that yeast expressing RNase H2 exclusively during G2 (the 'growth phase' of the cell cycle following DNA replication) were resistant to MMS, whereas yeast expressing RNase H2 only during S phase (the DNA replication phase) were more sensitive. This suggested that RNase H2 primarily acts to process R-loops during G2. In contrast, yeast expressing RNase H1 in either G2 or S phase were both able to survive in MMS. Surprisingly, RNase H2 expression in S phase actually induced more DNA damage, which required a special type of DNA repair called homologous recombination to fix. This pathway was not previously known to act during S phase. Therefore, this study may have revealed an unexplored repair pathway which counters damage caused by RNase H2 activity during DNA replication.
These results may explain why cells have evolved two different RNAse H enzymes. Luke clarifies: "We think that RNase H2 is the 'housekeeping' enzyme that repairs the majority of RNA-DNA hybrids, but it is strictly regulated by cell cycling and acts only in G2 phase, or after DNA replication." Luke and Ulrich speculate this may be because the additional RER activity of RNase H2 creates nicks in the DNA, which are a risk for double-strand breaks during DNA replication in S phase. Therefore, cells may have also evolved a secondary enzyme, RNase H1, which does not have RER activity and can act in all phases of the cell cycle, including S phase.
These findings help us to further understand how cells repair DNA damage associated with RNA-DNA hybrids and how impairment of this process contributes to disease.
Safety Precautions in the laboratory in the time of Covid-19
- Limit close contact – Distance from one another must be observed at all times. Movement should also be limited to essential trips.
- If possible, work remotely or stagger shifts to minimize the number of people in the lab.
- Observe proper hand hygiene at all times. There should be a hand washing station and a hand sanitizing area.
- Frequently touched surfaces must be kept clean and sanitized using products that meet the criteria set by the Environmental Protection Agency. It includes laboratory equipment and cabinet handles. Make sure to wear personal protective equipment while cleaning and sanitizing the lab.
- If someone in the lab is positive of Covid-19, the protocols established by the Center for Disease Control should be followed. Track the areas used by the infected person and clean and decontaminate it using EPA-recommended disinfectant.
Decontamination Solution (v1.4)
10% Store bought bleach (2L per 20L)
1% NaOH (200g per 20L)
1% Sparkleen or similar powdered detergent (200g per 20L)
Instructions for use:
- For most applications (Wiping down countertops, equipment, pipettes) the decon solution can be diluted 2-3X, wiped on, allowed to soak for several minutes and then rinsed off with distilled water and towels. More stubborn messes can be hit with undiluted mix.
- Glass and parts can be set to soak in straight or diluted (2-3X) decon mix, then washed as normal and rinsed with distilled water.
- Don’t let the decon mix come in contact with anodized aluminum, it will take the color right off!
- Latest, easiest decon mix. Dropped the addition of sodium bicarb since sparkleen contains a good bit of it and it provides the detergent for the wetting action. Thank you to the reader who suggested it!
Decontamination Solution (v1.3) (Old, legacy version)
10-15% Store bought bleach (100-150 mL/L)
1% NaOH (10 g/L)
1% Alconox/Sparkleen/dish soap (10 g/L) *
90 mM sodium bicarbonate (7.5 g/L) **
* Commercial versions use SDS, but at higher concentrations (=>1%) the SDS will tend to crash out. Unless you have the 2141-BG fragrance/emulsifier, either use a lower concentration of SDS (<0.1%) or use the above detergents *
** Sparkleen and Alconox have sodium bicarbonate already in it in high concentrations, up to
40% for Alconox, so the addition of bicarbonate may not be necessary.
Assuming Sparkleen has 30% bicarbonate, 10 grams of sparkleen has 3 grams bicarbonate, which would make a final solution that has 36 mM bicarbonate, which could still provide corrosion inhibition, depends how strongly you want to believe the 90 mM from the DNAzap patent. **
*** This decon mix will corrode aluminum and iron/cheap stainless steel at high concentrations and when treating for long periods of time. Good quality stainless hold up fine. ***
**** This mix is awesome for cleaning glassware, let a beaker sit in 0.5X or 1X decon mix for a while, the longer the better. It will sparkle after you rinse it! The high NaOH content is reminiscent of base baths used by chemists to etch a nice clean layer on their glass. ****
For more background on how I arrived at this recipe check out my second post on the topic, Just bleach it
Is autoclaving enough to get rid of RNAses from pipette tips and eppendorf tubes?
I'm consistently getting negative results with RNA extractions and I suspect RNAse contamination. Our workspace is very clean, but I am now concerned about our tubes and tips. Is autoclaving enough to eliminate RNAses from these, or are there any additional methods to keep them RNAse free?
No. Order RNAse-free pipette tips and tubes. Clean everything, including your gloves, with RNAseZap, before you reach into the baggies to get tubes/tips. RNAse zap on the pipettes, too. Don't even breath on the tubes/tips. Keep them in a special RNAse-free area when not in use. RNAse is on everything. Use DEPC-treated water only, too.
EDIT - Use filtered tips, too. I guarantee that you are spraying RNAse into your sample if you're not.
We buy RNase/DNase-free tubes and still autoclave them. They get poured from the big bag into containers and autoclaved. Filter tips are also RNase/DNase free but don't get autoclaved.
We use Qiagen kits to extract RNA. The only tubes and water used in this protocol come with the kit so you don't have to worry about making sure your own tubes are RNase free or DEPC treating your water (at least not for the extraction. if you're doing downstream experiments
The only thing I RNaseZap before extracting RNA is the rotor-stator tip that's actually going into the tube with the tissue in it. I know some people spray down their desk, their gloves, have a special RNA bench, etc. We don't have any of that and don't have problems.
This. Worked with RNA for the past 2 years. Required all new tips pretty often, with filters, special pipettes and a special area just for handling RNA. Everything (pipettes, bench, forceps etc) every 2 weeks. Added RNase inhibitors to anything that was being stored for any length of time. DEPC-treated water is also nice, but since DEPC is kinda dangerous we saved it for the most important steps.
Also, never autoclave the filtered tips. Once they are used toss em.
interesting, our group works almost exclusively with RNA, We have separate pipettes for RNA and other work, but exclusively use autoclaved tips (no filter) and microfuge tubes. Autoclaved water has also never proven to be a problem. We get great yields of intact RNA.
And I now know the newest addition to the wall behind the qPCR machine.
I've found that residual ethanol or phenol is the most common cause of poor RNA yields.
I agree. RNase seems to be a boogeyman and over the years I've gradually stepped down my anti-RNase precautions to the point where I don't even think twice when switching from standard DNA-related molecular biology work to RNA work. I've never observed any kind of mysterious sample degradation. I use water that comes straight out of a MilliQ type system and regular tips, no special pipets, etc.
I'm no longer working in a research lab, but I remember that some people actually autoclaved their RNase in order to sterilise it and it was still working afterwards. So I don't think that method is useful to actually eliminate RNases. There are some products to decontaminate the workspace althought it doesn't look like it is your problem.
One of our German post-docs once told me that no, some RNases can survive autoclaving (which blows my mind). I work in a RNA lab and it's disgustingly dirty, but we just use good quality tubes/tips and give a spray of RNase away here and there. We've never had any contamination issues.
If you're still having problems just toss everything and start fresh.
People are talking a lot about the external sources of RNAse contamination, but that is miniscule compared to the RNAses already present in the tissue you are extracting from. How are you extracting the RNA? there are several methods, my favourite is freezing as soon as possible in liquid nitrogen, and grinding up into a fine paste (keeping below freezing with liquid N) and mixing the paste with guanadinium thiocyanate/phenol extraction buffer (i.e. something like TriZol).
If you're not using a method similar, what does your extraction buffer contain to inhibit/denature the RNAses? the two buffers I use regularly contain either SDS or phenol - phenol being my favourite.
tldr: Your biggest source of RNAses is not you, your bench or your equipment when working from a source of RNA (unless you're transcribing it yourself of course). The biggest source of RNAses is the tissue/animal from which you are extracting RNA.
Research that is too dangerous?
However, the principle underlying gain-of-function research has been widely challenged over the past decade.
A classic and oft-cited example, which concerns many scientists, is Ron Fouchier and Yoshihiro Kawaoka’s research on the highly dangerous avian H5N1 influenza virus. Using a technique that passed the virus from one ferret to another many times over, these researchers were able to create an H5N1 influenza virus that could be transmitted to the species by aerosols.
The study was widely debated and the research was eventually put on hold. The U.S. government even urged scientific journals not to publish its full results, arguing that the information could be used by bioterrorists. The research was resumed in 2013.
Gain-of-function research has the potential to help prevent animal-to-human transmission of a virus with pandemic potential. However, this type of research must be carried out in highly secure laboratory, such as those known as BSL-4.
A view of the BSL-4 laboratory inside the Wuhan Institute of Virology after a visit by the WHO team, Feb. 3, 2021. (AP Photo/Ng Han Guan)
These laboratories are built to protect staff and researchers from becoming infected, and to prevent organisms from escaping. However, documents from U.S. Embassy officials have revealed that the biosafety standards at the BSL-4 laboratory at the WIV were not sufficiently rigorous. In addition, a number of researchers have suggested that the institute’s gain-of-function studies on bat coronaviruses were risky and might be harmful to humans if it escaped.
Did COVID-19 Really Escape From A Lab? The Evidence Suggests It Did
Wisconsin Congressman Mike Gallagher isn’t one for conspiracy theories. As a U.S. Marine Corps intelligence officer, he was trained to deal only in facts. And now the facts are leading him unmistakably to a conclusion that for more than a year has been dismissed as crackpottery.
COVID-19, he is convinced, escaped from a lab.
“Don’t mind me as I put on my tin foil hat,” he joked Tuesday before a radio interview in which he outlined the convincing evidence that the SARS-Cov-2 virus originated in the Wuhan Institute of Virology.
“Flash back about a year ago and change, when there were a few of us who were suggesting that it was a remarkable coincidence that there was the [biosafety] level-4 biosafety lab located where this whole outbreak started,” he said.
This lab also just happened to be the epicenter of the entire nation’s study of bat viruses. It is the home of Dr. Shi Zhengli, known as China’s “Bat Lady” because of her research into bat-borne illnesses such as the SARS-CoV-1 virus that caused Asia’s deadly SARS outbreak in 2002.
Although the Chinese government has sealed her files, it was known that she had collected bats from the Yunnan province that carry the closest known relative of SARS-CoV-2. This is significant, because Yunnan is 1,000 miles away from Wuhan, making it all but impossible for a bat in the wild in Yunnan to infect a human in Wuhan and start a pandemic there.
Early theories about the virus jumping to humans after an unlucky person ate a bowl of bat soup in the Huanan Seafood Wholesale Market in Wuhan were dismissed last May, effectively ruling out the market as the source of the outbreak. Moreover, bats hibernate in the winter months when the virus was supposedly first detected, so a human encounter with a wild bat was always a rather unlikely cause.
“In order to prove the wet market theory, you have to prove that the host species somehow traveled 1,000 miles but didn’t start an outbreak along the way,” Gallagher explained, adding that a far more probable origin was a virus that had escaped during “gain-of-function” research at the Wuhan Institute of Virology.
Gain-of-function is a type of medical experimentation in which viruses are induced to mutate into more transmissible and more dangerous versions. The goal is to study these mutations in order to better prepare humanity for the next deadly pandemic. Dr. Zhengling’s work, for instance, was focused on mutating the SARS-CoV-1 virus so as to prevent another deadly SARS outbreak.
This, Gallagher admits, is where things start to get a little crazy, especially since that research just might have been funded by a grant from the United States. Specifically the National Institutes of Health. Specifically the National Institute of Allergy and Infectious Diseases (NIAID). Specifically its director, Dr. Anthony Fauci.
Gallagher does not suggest that Dr. Fauci and the Chinese government were conspiring to unleash a weaponized virus on the world, but rather that American researchers—who were prevented from doing gain-of-function research under a 2014 Obama Administration dictate—outsourced the work to the Chinese.
In 2011, Dr. Fauci co-wrote a Washington Post op-ed entitled “A Flu Virus Risk Worth Taking,” in which he argued that “important information and insights can come from generating a potentially dangerous virus in the laboratory.”
In other words, a decade ago he strongly supported gain-of-function research.
“Understanding the biology of influenza virus transmission has implications for outbreak prediction, prevention and treatment,” he wrote. “In defining the mutations required for mammalian transmission, public health officials are provided with genetic signatures that, like fingerprints, could help scientists more readily identify newly emergent, potentially harmful viruses, track their spread and detect threatening outbreaks.
“The ability to identify such viruses even a few months faster than by conventional surveillance provides critical time to slow or stop an outbreak.”
In 2014, however, President Obama’s White House put a three-year moratorium on gain-of-function research into SARS, MERS, and influenza viruses. That same year, Dr. Fauci’s NIAID awarded a $3.4 million grant to EcoHealth Alliance, a group that seeks to protect humanity from viruses that might jump from other species.
EcoHealth Alliance, under the direction of Dr. Peter Daszak, used part of its NIAID grant on a project entitled “Understanding the Risk of Bat Coronavirus Emergence.”
“Most emerging human viruses come from wildlife, and these represent a significant threat to public health and biosecurity in the US and globally, as was demonstrated by the SARS coronavirus pandemic of 2002-03,” he wrote in the project’s Public Health Relevance Statement. “This project seeks to understand what factors allow coronaviruses, including close relatives to SARS, to evolve and jump into the human population by studying viral diversity in their animal reservoirs (bats), surveying people that live in high-risk communities in China for evidence of bat-coronavirus infection, and conducting laboratory experiments to analyze and predict which newly-discovered viruses pose the greatest threat to human health.”
Since gain-of-function research was effectively banned in the United States, EcoHealth Alliance contracted with the Wuhan Institute of Virology in Wuhan to study coronaviruses found in bats in the Yunnan province…the very same bats that have the strain of coronavirus that is closest to SARS-CoV-2.
The Wuhan Institute of Virology vehemently denies performing any gain-of-function experiments, but since Dr. Zhengli’s files are sealed, it is impossible to know exactly how she was spending EcoHealth Alliance’s grant money.
Dr. Zhengli herself has come under fire for “unsafe laboratory practices” that may have allowed SARS-CoV-2 to escape. As far back as 2018, a cable from the U.S. Embassy in Beijing warned that “during interactions with scientists at the Wuhan Institute of Virology laboratory, they noted the new lab has a serious shortage of appropriately trained technicians and investigators needed to safely operate this high-containment laboratory.”
Dr. Zhengling has admitted that she did not perform coronavirus research in biosafety level-4 areas that require hazardous materials suits and extreme precautions. Instead, she said she worked in a biosafety level-2 lab, which requires only moderate precautions.
On balance, which seems more likely: That a virus escaped from a lab that was performing experiments in known unsafe conditions or that a bat somehow traveled 1,000 miles from the caves of Yunnan to the Wuhan wet market and bit someone?
All Gallagher wants is a further investigation of what is looking likelier and likelier by the day—that the virus escaped from the lab and the Chinese government is trying to cover up its embarrassing slip-up… just like it tries to cover up all of its embarrassing slip-ups.
This isn’t so much a conspiracy theory as it is a rational look at the available evidence and when the evidence is all pointing in one direction, it’s time for investigators to head there.
#1 – Schedule cleaning time
Working in the lab can be hectic. While scheduling cleaning can be inconvenient, all your hard work will be for naught if samples keep getting contaminated because the lab is dirty. To make the most of your time, you can plan lab clean-up hours using a laboratory information management system (LIMS). A cloud-based LIMS can efficiently manage many aspects of your lab, including workflow, inventory, and results analysis it can also regularly schedule hours for cleaning far in advance, allowing you to assess your lab’s cleaning requirements and assign members of the lab to specific cleaning-related tasks as needed.
The main reasons why eating and drinking are not permitted in areas using or storing hazardous materials are personal safety risks and risks of non-compliance with regulatory or granting agency requirements that may impact an individual, a work unit, or the institution as a whole.
Personal Safety Risks
Personal safety risks can result from cross-contamination and ingestion. Contamination can result from contact with contaminated gloves/hands, airborne materials settling out or condensing on surfaces or utensils, or placing consumable items on a contaminated surface.
Prudent Practices in the Laboratory by the National Research Council includes these precautions for minimizing exposure:
- Eating, drinking, smoking, gum chewing, applying cosmetics, and taking medicine in laboratories where hazardous materials are used should be strictly prohibited.
- Food, beverages, cups, and other drinking and eating utensils should not be stored in areas where hazardous materials are handled or stored.
- Glassware use for laboratory operations should never be used to prepare or consume food or beverages.
- Laboratory refrigerators, ice chests, cold rooms, ovens, and so forth should not be used for food storage or preparation.
- Laboratory water sources and deionized laboratory water should not be used as drinking water.
- Laboratory chemicals should never be tasted.
- A pipette bulb or aspirator should be used to pipette chemicals or to start a siphon pipetting should never be done by mouth.
- Hands should be washed with soap and water immediately after working with any laboratory material, even if gloves have been worn.
The main compliance and grant risks summarized below include regulations (either federal or state laws), consensus standards, and granting agency requirements. These mandates are also included
in University programs for occupation health and safety and radiation protection.
The OSHA Lab Standard and the University Chemical Hygiene Plan prohibit eating/drinking in areas where hazardous chemicals are in use.
The OSHA Bloodborne Pathogens Standard and the University Exposure Control Program prohibit eating/drinking in areas where a reasonable likelihood exists for exposure to blood or other
potentially infectious materials.
The University Biological Safety Program states that eating, drinking, smoking, handling contact lenses, or applying cosmetics are not permitted where rDNA research is done, or where there is
reasonable likelihood of exposure to potentially infectious material. This is based on National Institutes of Health Guidelines for Research Involving Recombinant or Synthetic Nucleic Acid Molecules and on Biosafety in Microbiological and Biomedical Laboratories.
The Iowa Department of Public Health, Bureau of Radiological Health and the University Radiation Protection Guide prohibits eating/drinking in areas where radioactive materials are present.
EPA regulations focus mainly on materials management and environmental impacts. However, regulatory inspectors may refer issues regulated by another agency to that agency. Therefore, EPA
may refer occupational issues to OSHA.
Granting agencies such as the National Institutes of Health and the Department of Defense require that both the unit receiving the grant and the institution as a whole to be in compliance with their
guidelines and the regulations of other agencies such as OSHA, EPA, and IDPH.
Based on the information cited above, it is the University’s policy that eating and drinking are not permitted in areas where chemical, radiological, and/or biological materials are used or stored.
RNase AWAY™ surface decontaminant
Thermo Scientific&trade RNase AWAY&trade surface decontaminant eliminate RNase and DNA from laboratory surfaces. It is ideal for decontaminating apparatus, bench-tops, glassware and plasticware, and it reduces dependency on carcinogenic DEPC treatments and saves time needed to bake glassware.
- Use on pipettes, gel boxes or RNA or DNA prep areas
- Leaves no residue to interfere with gel polymerization or staining
- Chemically stable and nonabrasive
- Contains no strong acids
|7000 - MBP||RNase AWAY, 250 ml|
|7002 - MBP||RNase AWAY, 475 ml spray bottle|
|7003 - MBP||RNase AWAY, 1 liter|
|7005-11 - MBP||RNase AWAY, 4 liter|
Prices and ordering
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