Hi all,
I am developing a high-throughput RNA extraction protocol for xylem vessels. Pre-emptively, the samples are going to be homogenized in a Genogrinder with a cryoblock attachment and then transferred to a 96-well format for total RNA extraction. For ease of transfer to the 96-well, I was thinking of maybe mixing the homogenized tissue with RNAlater-ICE purely for the fact that transfer of dry material to a 96-well will be too messy and not very high-throughput. Does anyone have experience with RNAlater altering the transcriptome gene expression profile? I read these attached articles on normal RNAlater and the influence it can have, but they only submerged whole tissues instead of ground tissues. What would be a good alternative to RNALater and RNALater-ICE? I am still inbetween using a 96-well plant RNA extraction kit vs CTAB RNA extraction. Would submerging the homogenized xylem in for eg. a kit's lysis buffer greatly affect the RNA integrity? Thanks in advance! Article Transcriptome and proteome responses in RNAlater preserved t...
Article Nonrandom RNAseq gene expression associated with RNAlater an...
Hi
RNA later is generally used when the whole the integrity of the RNA inside the tissue needs to be maintained, if it is not possible to freeze in the Liquid Nitrogen, immediately. It is possible that RNA later may also cause problem in RNA isolation, if not removed properly. Its removal is relatively easy from the surface of whole tissues, than if it is added to the homogenized tissue. For your experimental needs, the TRIZOL/TRI reagent may be a better good option. It maintains the quality of RNA in the homogenized tissue. And it should be useful of high-throughput isolation of RNA
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