I want to use QRTPCR to measure prepro EGF levels in the mouse kidney. I have not been able to find papers that provide sequences for Sybr primers. Most papers only do RT-PCR, or dot blots.
You can do a couple things. One is that you design your primers using the sequence available in the database (GenBank, etc), which is what we do in my lab. There are different free primer designing programs available on-line that you can use to design the primers you wish. The major issue with this is that you need to know where the exon-intron junctions are within the mRNA so that either one of the primers or the probe will sit at the junction and only amplify the cDNA and not not genomic DNA. Given that you are working with mice, you can buy commercially available primer sets from the biotech companies. Although it may be a tad expensive than home-made primers, pre-made primers are usually optimized for the specific qPCR reagent so you don't have to spend too much time optimizing the assay. Sorry for a long answer. Hope this helps.
Hello
I agree with Yasuhiro Kobayashi , Kindly look at the link .
http://citeseerx.ist.psu.edu/viewdoc/download?doi=10.1.1.948.4218&rep=rep1&type=pdf
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