No RNA after Trizol extraction
Posted 02 December 2009 - 02:40 AM
Posted 02 December 2009 - 05:11 AM
I am not sure, how much of RNA to expect from Mango fruit pulp, but I guess it should not be much...its quite possible that you are actually getting the polysacharides from the solution. I once tried isolating RNA from banana leaves and always used to end up with perfect looking huge pellet, but this never yielded to dissolve completely and my quantification were grossly wrong possibly because of the huge amounts of polysacharides in the leaves...Try adding more of chloroform maybe 1ml: 300- 350ul against 1ml:200 ul..This might help
Posted 08 December 2009 - 10:57 PM
Posted 09 December 2009 - 01:31 AM
Trizol might not work for every plants..it didnt work for banana in my case, but worked for leaves and flowers of maize, sunflower, rice, barley, sorghum and even watermelon fruit...
the brown/greenish color that you see is normal..the color remains even in the pellet sometimes, but i've used it for microarray experiments successfully
Posted 09 December 2009 - 01:45 AM
Posted 09 December 2009 - 02:36 AM
Did u try with some other method already??
Edited by gogreen, 09 December 2009 - 02:43 AM.
Posted 14 December 2009 - 06:19 AM
Posted 14 December 2009 - 09:47 AM
Posted 21 December 2009 - 11:59 PM
75 mg tissue + 0.75ml extraction buffer
[100 mM Tris-Cl (pH 8.2),1.5 M NaCl, 30 mM EDTA (pH 8.0), 2% CTAB and b-mercaptoethanol]
Incubated 65C for 20 min--cooled + 0.5ml trizol
Rest everything by Trizol protocol with inclusion of additional chloroform wash before isopropanol precipitation..
Got 300ng/ul RNA in the volume of 50 ul. There was very small genomic DNA contamination that could be removed by DNase treatment
Edited by ram, 21 December 2009 - 11:59 PM.