In order to determine the effects of endocrine perturbators in RAT cells from colon tumors, I want to determine their impact on mRNA expression of esr 1 and 2 . Due to the low level of esr 1 expression in the rat colon, I need to use validated primers.
Does anyone have primer sequences for esr or the address of companies which sell these products?
If you are amplifying the coding region there is a good chance that the sequence is conserved in mouse. In that case you can use the NCBI qPrimer depot (http://mouseprimerdepot.nci.nih.gov/).
Thank you for your suggestion but I did not ''found'' equivalence between mouse and rat sequence for esr.
After some trials with this tool, it sounds interesting but as you mentionned optimisation is required. Due to my ''urgent need'', a ''all-in-one solution'' (design+validation) sounds more appropriate.
Nevertheless, I'eve noticed the roche ologo probe link for further investigations.
thank you for your help
Pay attention to estrogen receptor splice variants that are expressed along with the full length transcript (PMID: 7743517).
Hi, I always check the roche oligo probe library
https://www.roche-applied-science.com/sis/rtpcr/upl/index.jsp?id=uplct_030000
or the Qiagen website. 99% of the time the primers recommended worked for me.
However, like I had to do for a couple, you should alwaystest and further optimize your PCR / qPCR with a less important RNA (also include a standard curve if doing quantitaion. good luck
Thank you for your answer which is in accordance with that of Dr Moser.
I'd like to thanks too Dr Pfeffer for its pertinent remark on splice variants.
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