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elader

Member Since 14 Jun 2012
Offline Last Active Oct 15 2012 09:25 AM
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Posts I've Made

In Topic: Genomic DNA contamination of RNA

14 June 2012 - 04:17 AM

You can never get rid of all genomic DNA contamination. One way to measure it if you are anal retentive about it is to have a pair of primers that both lie in the same exon - do a real time experiment with/without RT in the cDNA synthesis step - theoretically you shuold get no signal from the no-RT control. The delta Ct between the no-RT and +RT will tell you the % genomic DNA (of course, there is variation in the RT efficiency), but you'll still get the idea.

In Topic: My RNA extraction failed (From a newbie )

14 June 2012 - 04:13 AM

You cant store undisrupted tissue in Trizol, the RNA will degrade. Either homogenize it in trizol and store it or snap freeze it or store it in RNAlater - dropping a piece of tissue in trizol and NOT immediately disrupting it is a very bad thing - slow lysis and release of nucleases. That is absolutely your problem. If you are having lengthy dissections and accumulating sample, RNAlater is probably the safest way.

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