I need to extract RNA mycelium of fungi from top agar medium, using Trizol or PIG B reagent. Everything is fine except in the final step, i still get the agar persisting in my RNA sample. The agar mixes with RNA sample and it becomes hard to go into cDNA synthesis. How do I remove the agar?
Dear Sandesh,
try heating the TRIzol with your mycelia at 65 degrees for 10 mins (water bath). It will not affect your RNA. Briefly vortex and allow it to cool at RT. then incubate the tubes at 2-8 degrees for 30 mins. agar will be solidified and carefully aspirate the trizol mix into a fresh tube and follow the subsequent steps. Try a couple of samples first.
Best of luck.. will be happy to know if this works.
SD
I tried but it didnt work. The conc was same or RNA. and the gel persisted. No change at all. Can you suggest any other ways?
I tried the RNA concentrater columns which is to be used after getting the supernatent state . It gave no agar solidification , however , the elution acccording to protocal is only 15ul, which is too less for my samples as well as the concentration of RNA is 10 times less.
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