Can anyone help me with an issue:
I usually do total RNA extraction with the CTAB protocol and using LiCl to precipitate the RNA ... At this stage of precipitation, the protocol recommends I place the LiCl and stock overnight at 4 ° C ... But I can leave the - 80 ° C for hours, let's say let's advance precipitation? I did the extraction leaving at -80 ° C for two hours and then following I realized that no pellet was formed, so when I finished the extraction no RNA and I went to quantify, there was nothing RNA ... Is the error was precisely in the stage of precipitation, I would have expected more even putting in a freezer at -80 ° C?
LiCl precipitation is not especially affected by temperature but is very sensitive to centrifugation speed and time. What spin conditions are you using please? You may find the attached taken from the thermos fisher website of interest
The highest centrifugation step is one 30 min, the others are all 10 min ... But today I realized that leaving at 4 ° C overnight the samples get better quantification and higher quality than at -80 ° C overnight. .. So maybe the precipitation temperature, at least in plant, can influence something ...
Thanks for the material, I'll pay attention to the things he says ... Thanks Paul Rutland!
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