Comments on: RNA Isolation – Jake’s O. lurida Ctenidia Control from 20150422 http://onsnetwork.org/kubu4/2015/05/07/rna-isolation-jakes-o-lurida-ctenidia-control-from-20150422/ University of Washington - Fishery Sciences - Roberts Lab Tue, 09 Apr 2019 18:37:58 +0000 hourly 1 http://wordpress.org/?v=4.0 By: Jay Dimond http://onsnetwork.org/kubu4/2015/05/07/rna-isolation-jakes-o-lurida-ctenidia-control-from-20150422/#comment-1475 Thu, 16 Jul 2015 20:34:59 +0000 http://onsnetwork.org/kubu4/?p=1325#comment-1475 OK – thanks for the clarification. Good to known that brown is ok. Corals have tons of pigments, so normally I wouldn’t be too surprised, except that the protocol talks about a clear or white film.

]]> By: kubu4 http://onsnetwork.org/kubu4/2015/05/07/rna-isolation-jakes-o-lurida-ctenidia-control-from-20150422/#comment-1474 Thu, 16 Jul 2015 20:25:00 +0000 http://onsnetwork.org/kubu4/?p=1325#comment-1474

Just for clarification, the image in this post is of a chunk of tissue prior to homogenization, not a RNA pellet.

However, it’s not uncommon to have “discolored” pellets. Pellet discoloration is heavily influenced by the source tissue. There are going to be molecules (e.g. various polysaccharides) that will get carried through the extraction and contribute an “off” color to the pellet. Some tissues (e.g. digestive gland from marine molluscs) always produce pellets that are brown.

In the end, the discoloration doesn’t seem to have any noticeable impact on RNA quality (260/230, 260/280 and Bioanalyzer traces), nor on downstream applications.

]]> By: Jay Dimond http://onsnetwork.org/kubu4/2015/05/07/rna-isolation-jakes-o-lurida-ctenidia-control-from-20150422/#comment-1473 Thu, 16 Jul 2015 17:15:50 +0000 http://onsnetwork.org/kubu4/?p=1325#comment-1473 My RNA pellet from the coral samples looked similar. I thought the pellet would be clear – why the brown color?

]]> By: DNase Treatment – Jake’s O.lurida Ctenidia RNA (Controls) from 20150507 | Sam's Notebook http://onsnetwork.org/kubu4/2015/05/07/rna-isolation-jakes-o-lurida-ctenidia-control-from-20150422/#comment-699 Thu, 14 May 2015 23:43:04 +0000 http://onsnetwork.org/kubu4/?p=1325#comment-699 […] the O.lurida RNA I isolated on 20150507 showed residual gDNA via qPCR, I treated 5μg of RNA from each sample using the Turbo DNA-free Kit […]

]]> By: qPCR – Jake O.lurida ctenidia RNA (Control Samples) From 20150507 | Sam's Notebook http://onsnetwork.org/kubu4/2015/05/07/rna-isolation-jakes-o-lurida-ctenidia-control-from-20150422/#comment-681 Wed, 13 May 2015 00:28:31 +0000 http://onsnetwork.org/kubu4/?p=1325#comment-681 […] qPCRs on the O.lurida total RNA I isolated on 20150507 to assess presence of gDNA carryover with Oly Actin primers (SR IDs: 1505, […]

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