I am planning to characterise the transcriptome of pre-anthesis kiwifruit anthers. The samples are being harvested on my behalf and sent down by post, and I cannot ask the harvesters to dissect out the anthers themselves. This means the samples will need to be preserved for up to two days.

I am planning to isolate the RNA through bead-beating and a trizol extraction.

My proposed solution is to harvest the whole young buds, and place them in RNALater. Unfortunately, the fatty and hairy surface of the kiwifruit buds mean there is poor penetrance of the RNALater solution to the organs of interest (the anthers). I have tried vacuum infiltration, to little success.

Another solution I have tried is to add a non-ionic detergent such as Triton-X-100 or Tween-20 to the RNALater solution. This appears to dramatically increase penetrance (the tissue inside appears greener, and I am able to produce a band on an endpoint RT-PCR gel. Unfortunately there is little published information on use of non-ionic detergents in RNALater solutions.

I was hoping to test the effects of this combination by visually assessing the RNA integrity through a simple gel analysis of the rRNA bands of the total RNA, but there is not enough in a sample to do this. So too is there not enough to do a Qubit RNA IQ assay.

My question is: Am I introducing any potential loss of RNA integrity by combining the RNALater solution with a non-ionic detergent?