a protein contain a histidin tag will be selectively bound to metal-charged media such as Ni Sepharose High performance (HP) and Ni Sepharose 6 Fast Flow (FF) while most others cellular proteins will not bind weakly.

Search the literature first using PubMed for published primers. We used primer sets including those for mouse Tnfa and IL17a genes that were published previously by others and validated in our lab (references are given in our paper: J. Immunol. 2010; 184:3008). One can also design primer sets using several sources, including IDT. You can blast the primer sequences to make sure that the primers recognize the specific genes and not somewhere in 99% of non-coding genome. Regardless of the source, you must carefully choose the primer sets and importantly validate them according to MIQE guidelines. These include melt curve analysis and exclusion of primer-dimer (check on agarose gel). Validation of primer sets is an absolute requirement and for some genes it will be difficult. There are several instances in the literature in which the investigators failed to validate their primer sets. This creates uncertainty and huge problems for others who subsequently wish to use these primers. Being qRT-PCR is the gold standard for gene expression and SYBR Green method is economical and preferable to use, spending some time on careful validation will go a long way.

Check out http://primerdepot.nci.nih.gov/ and Roche Applied science website (https://www.roche-applied-science.com/sis/rtpcr/upl/index.jsp).

One can also design primers by accessing IDT and Applied Biosciences websites, among others.