If we used Milli Q with BioPak purification water in place of DEPC, can it work or it will affect the RNA purity ?
MilliQ is enough as long as the machine is properly maintained in terms of sanitization/sterilization and as long as the tubes and glassware you are using are clean!!!
Sometimes milliQ quality is good enough for RNA as well. You may try both ways and see what gives you better result for the large-scale long-term experiments.
Another thing people are using is RNasin (RNase Inhibitor). When stock was low I have tried without it and RNA still stable. One just needs to be very careful. But generally DEPC and RNasin are good addition to RNA experiments.
DEPC is good for RNA related work, otherwise you can use Milli Q from water purification systems attached such as DNae/RNase filter membrane.
MilliQ is enough as long as the machine is properly maintained in terms of sanitization/sterilization and as long as the tubes and glassware you are using are clean!!!
Some kits (RNeasy Mini-kit QIAGEN, for example) recommend not using DEPC water. MilliQ is more than enough, but with certain cares like said above.
We are using milliQ. We just sterilise it in autoclave for an hour at 120º if it's for RNA.
Also it depends on the downstream applications of your RNA because DEPC inhibits many sensitive reactions for example DEPC inhibits transcriptase. But better to treat all your glassware with DEPC and bake them to make RNase free.
If you are using any kits for RNA isolation, normally they include some RNase inhibitor in one of the components.
Use RNase inhibitors like RNasin for reactions involving RNA, and DEPC treated sterile water to rinse equipments. That is what we practice in our lab. When water is autoclaved after treatment with DEPC, it is completely dissociated, and the DEPC treated water is safe.
So, are you saying DEPC-containing water or DEPC-treated water?
http://www.lifetechnologies.com/ca/en/home/references/ambion-tech-support/nuclease-enzymes/tech-notes/rnase-and-depc-treatment.html
I hope the link could help.
By my own experience using MilliQ water is more than enough for RNA analysis if the apparatus is properly maintain. Also DEPC treated water is slightly acidic and it gives lower than expected 260/280 ratio, and this might lead to confusion.
I was working with very low amount of tissue and usage DEPC gave me far better results (amount and quality of mRNA (RIN))
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Molecular Biological Techniques