Hello everyone,
Since few time we are isolating RNA by Sigma Trizol method from root and shoot of our rice sample and still we are confusing that why we are getting much intense band of while loading only 1 micro liter sample of RNA in 1.5% agarose gel.
The 260/280 ratio we are getting 1.9 approx for RNA.
Can anyone explain the reason behind it, if there is any solution to improve our RNA quality please suggest us.
Kindly find the attached file.
Hello, In my humble-not so experienced opinion, there is still genomic DNA in your extraction results, you need to perform a DNAse Step.
also, the lower band could mean that there is degradation of RNA or degradation of DNA, you should perform DNAse treatment and see what happens after this...
if there is infact degraded RNA, the RNA trizol sigma web page gives you some troubleshooting guides:
If there is degradation of the RNA:
the tissues may not have been immediately processed or frozen after removing from the animal.
the samples used for isolation or the isolated RNA preparations may have been stored at –20 °C instead of –70 °C as specified in the procedure.
cells may have been dispersed by trypsin digestion.
aqueous solutions or tubes used for procedure may not have been RNAse‑free.
formaldehyde used for the agarose gel electrophoresis may have had a pH value
- Badges
- Science topic
- Similar topics
- Agriculture
- Horticulture
- Seeds