Hello everybody ,
Can anybody guide me about the best gene for plants as control for expression profiling(qPCR).
i m working on Coleus forskohlii ABC transporter proteins
Dear Gulfam
Actin and Ubiquitin are universal house keeping genes. They will be most appropriate for qPCR in any plant species.
You can also use 18S as your housekeeping gene. It's better to use two housekeeping genes on your gene expression studies.
Hello,
which reference genes are stable enough depends on your experiment and varies from experiment to experiment even within the same tissue of the same species! You need to test several candidate reference genes every time you set up a new RT-qPCR experiment. The way to go is to make a list of minimum 10 candidate reference genes (e.g. from literature) and test all of them on a representative subset of your samples. Pick those that are stable enough.
Ribosomal RNA is not a good choice according to several authors since it is not always representative for the amount of mRNA in your extract.
Read papers and documents from Vandesompele and Hellemans from the Biogazelle company: https://www.qbaseplus.com/knowledge/blog/four-tips-rt-qpcr-data-normalization-using-reference-genes
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