Dear All,
I’m planning to take freshwater samples for microbial metatranscriptomics analysis (RNA-seq). The problem is, the journey from the sampling site to my lab might take >5 hours. I’m going to filter the samples into a sterivex filter on-site, and immediately extract the RNA once I reach the lab. What is the most convenient and appropriate method to preserve the sterivex filter during the journey?
i) RNAlater : Some researchers/papers reported on the use of RNAlater solution may compromise the transcriptome due to the osmotic cell response to the high external salt concentration. Hence, I may not use this method.
ii) Flash freezing with liquid nitrogen
iii) Flash freezing with dry ice + 100% ethanol bath
Has anyone tried preserving the sterivex filter with dry ice+ethanol bath? Between liquid nitrogen and dry ice+ethanol bath, which method is more convenient to be used in the field (safety, etc)? Any effects on the RNA or transcriptome profiles?
Thanks!
Methods may be appropriate. Another option; cold transport (0, 1, 2, 3 oC) by preparing different dilutions (-1, -2, -3, -4, -5, -6) with suitable medium (like as pepton water, aseptically prepared). 5 hours is not a very long time. So, you could transport them alive. So you can take it to the lab without using sterivex filter (and others). You can carry out the transportation with ice batteries in the transport box or with the car-cooler-refrigerator.
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