|Register||Search||Today's Posts||Mark Forums Read|
|PCR - Polymerase Chain Reaction Forum PCR - Polymerase Chain Reaction Forum. Discuss and ask questions about PCR troubleshooting, PCR protocols and methods, PCR products, and PCR theory.|
| ||LinkBack||Thread Tools||Display Modes|
Within-group trend of decreasing mRNA expression
Hi all! I have come across something weird in my latest qPCR study, and I would like to hear your input. I have exposed fish to a drug and have measured the expression of a couple of genes a a specific tissue. What I have found is that I have a strong trend of decreasing mRNA expression WITHIN my exposure groups, and that this trend coincides with the order in which total RNA was isolated from my samples. The lab assistant doing the RNA isolation did it non-randomized and in groups of six samples. 230/260 and 260/280 ratios were all above 2, gel electrophoresis showed no degradation and the total RNA concentration did not show any discernible trend. Could it be that the within-group trends I'm seeing is due to the fact that it takes a little longer to isolate RNA from the "later" samples (some thawing of tissue etc) and that some mRNA is degraded during this time? And I have to point out that the trends are in the raw expression data, and disappear when I normalize to the housekeeping gene (which also shows the trend).
So what do you think? Have you encountered anything like this before? And most importantly, does this affect the credibility of my data?
|decreasing , expression , mrna , trend , withingroup|
|Thread||Thread Starter||Forum||Replies||Last Post|
|Human Cytome Project - Update 24 Jan. 2005||Peter Van Osta||Cell Biology and Cell Culture||1||08-01-2010 02:18 PM|
|Human Cytome Project - an idea - Update 19 April 2005||Peter Van Osta||Cell Biology and Cell Culture||1||06-01-2009 02:17 PM|
|Variety Cultivar or Form, how do I know ?||Duncan||Botany Forum||13||10-28-2005 08:49 PM|
|Human Cytome Project - Update 6 Jan. 2005||Peter Van Osta||Cell Biology and Cell Culture||0||01-06-2005 10:18 AM|
|Sci.chem FAQ - Part 1 of 7||Bruce Hamilton||Chemistry Forum||0||01-15-2004 08:06 AM|