Hello all,
I am working with Mango pulp to isolate Total RNA. My lab lacks Spermidine which precipitates and removes DNA from sample during first stage in RNA isolation. I am following CTAB method for RNA isoloation from Pine leaves. Please confirm that can 3M Sodium Acetate be used as its substitute. And please give the working concentrations and names of other chemicals that can be used as Spermidine substitutes. Thankyou.
I am not familiar with performing isolation from pulps and plants, that being said, 3M Sodium acetate will not precipitate only DNA as I often use it to precipitate RNA. I would recommend using a phenol:chloroform or trizol extraction and then treating with DNase I to degrade the DNA and leave the RNA intact.
Hi, even using spermidin and LiCl precipitation there is some residual DNA to the precipitated RNA. Sodium acetate also will precipitate not only RNA but DNA as well. As Julie said the best way is to treat with DNAse I .
Good luck!
Acidic phenol:chloroform separation followed by DNase I treatment.
Junaid Kori,
I am using this same protocol and I have the same doubt. Did you have the chance to use DNase as a substitute for spermidine? Did it work?
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