Sam's Notebook » 06:50-10 http://onsnetwork.org/kubu4 University of Washington - Fishery Sciences - Roberts Lab Thu, 08 Nov 2018 21:47:12 +0000 en-US hourly 1 http://wordpress.org/?v=4.0 qPCR – DNased Abalone Dg RNA from 20090625 http://onsnetwork.org/kubu4/2009/07/10/qpcr-dnased-abalone-dg-rna-from-20090625/ http://onsnetwork.org/kubu4/2009/07/10/qpcr-dnased-abalone-dg-rna-from-20090625/#comments Fri, 10 Jul 2009 16:52:00 +0000 http://onsnetwork.org/kubu4/?p=957

Ran qPCR on gDNA (06:50-10) to test new primers (H.iris_actin_intron_Fw/Rv) designed to bind only to a region in an intron of the H.iris actin gene. Hopefully there’s enough homology between H.iris (primer source) and H.cracherodii (template source) for this to work. PCR setup/plate layout is here. Anneal temp 50C.

Results: No signal. :(

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qPCR – DNased Abalone Dg RNA from 20090625 http://onsnetwork.org/kubu4/2009/07/08/qpcr-dnased-abalone-dg-rna-from-20090625-2/ http://onsnetwork.org/kubu4/2009/07/08/qpcr-dnased-abalone-dg-rna-from-20090625-2/#comments Wed, 08 Jul 2009 22:22:58 +0000 http://onsnetwork.org/kubu4/?p=963

Ran qPCR on DNased Abalone Dg RNA (07:12 Set), gDNA (06:50-10) and clean cDNA (from 20090422) using primers (H.crach_h-1fg_intron_Fw/Rv) designed to bind only to a region in an intron of the H.cracherodii hemocyanin gene. PCR setup/plate layout is here. Anneal temp of 50C was used.

Results: No PCR products in any samples, not even the positive control. It seems that the primers don’t work. Will design new primers, probably from a different species of abalone since there was essentially only one gene in H.cracherodii that had any intron sequence available..

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