Whether anybody know the Primer sequences (quantitative RT-PCR analyses) in fetal Rattus norvegicus for deiodonases (D1,2 & 3), thyroid transporters (MCT8 & OATP1C1), thyroid receptors (α & β), TSH receptor, NIS, TTF1, TPO, TG and two reference genes for normalization?
Go to NCBI and get the sequence of genes of your interest. You can design your own primers using either NCBI primer design tool or Primer 3. The primer design depends on what you want to explore. There are many conditions for the primers to use in qPCR. Generally, primers with high GC content and amplicon size of less than 200 bp are preferred in qPCR.
For the thyroid samples, you can choose from the following reference genes:
1- hypoxanthine phosphoribosyl-transferase1
2- ribosomal protein LI3a (RPLIA)
3- beta-2-micro-globulin (B2M)
4- beta-actin (ACTB)
5- glyceraldehyde-3-phosphate dehydrogenase (THE BEST, according to the study of Chantawibul et al. J Med Assoc Thai. 2012 Mar;95 Suppl 3:S36-40).
Best wishes.
Ahmed
you can indeed design your own primers or else simply search for publications where these mRNAs have already been measured by qPCR. Authors normally provide a table or a reference for the primers they have used.
Veerle
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