I am using kits from Thermorscientific to extract total RNA. I extracted more than 20 samples and the results were various. Some of them had high concentration and the other ones were low. I do not know if it is because I processed too many samples at the same time. I use the same amount of sample for each reaction.
When I need to process a large number of RNA extractions in one day (have done 96), I do small-batch lysis (e.g. bead-milling mouse tissues) and then leave the samples in the SDS or Guanidium reagent until I finish lysing all the samples. I then do organic phase extraction or column purification in batches of 24. Importantly, make a note of which samples were processed in which batch, so you can check for batch effects later. Even more importantly, make sure each batch has mixed sample types. Do NOT process all the controls in one batch and all the treated samples in another!
Good luck!
Probably the problem is the instability of your sample. Try to reduce the working time with less samples and check.
The biggest factor in any extraction is lysis.. make sure that your samples are effectively mixed with the extraction buffer... if they are not mixed well you will not get good extraction...
When I need to process a large number of RNA extractions in one day (have done 96), I do small-batch lysis (e.g. bead-milling mouse tissues) and then leave the samples in the SDS or Guanidium reagent until I finish lysing all the samples. I then do organic phase extraction or column purification in batches of 24. Importantly, make a note of which samples were processed in which batch, so you can check for batch effects later. Even more importantly, make sure each batch has mixed sample types. Do NOT process all the controls in one batch and all the treated samples in another!
Good luck!
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