I have already done a RT-PCR with RNA from rabbit testis. I used Beta-2 microglobulin as housekeeping gene. Some problems were occured because the expression levels were very different among samples. Any suggestion for a new housekeeping to use?
Hello Francesco,
I stumbled across answers for a similar question:
https://www.researchgate.net/post/Which_housekeeping_gene_is_suitable_for_rabbit_tissue
You may find it useful to use a kit that helps you identify the optimal reference genes for your samples using Housekeeping Genes PCR Arrays in case of further difficulties:
https://www.qiagen.com/be/shop/pcr/primer-sets/housekeeping-genes/
Sincerely,
Check our article, we used ribosomal 28S subunit specific primers.
Article Transposon-Based Reporter Marking Provides Functional Eviden...
- Optimal housekeepers tend to be tissue/species specific
- Thus scour literature in the first instance
- Failing that identify housekeepers commonly used in rabbit studies
- Alternatively, use program scripts like Gene Norm which screen a panel of Housekeepers and identify the most expressed and stable in your system
- They would probably include the usual ubiquitous suspects
4. 18srRNA
Google universal housekeepers and you will obtain a list of genes including the above that could be screened in your system with programs like Gene Norm or Norm Finder
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