Glyceraldehyde-3-phosphate dehydrogenase ( GAPDH) is a common reference genes in relative RT-qPCR experiments 8. Furthermore, a groundbreaking analysis of RNA-seq data criticized the indiscriminate use of common reference genes 7. Although one advantage of using reference genes is that their expression does not change under different physiological and experimental conditions 3, 4, numerous announcements have prohibited the use of routinely used reference genes blindly 5, 6. Reference genes have been routinely used in gene expression analyses in traditional cancer studies 1, 2. As a result, choosing the appropriate reference gene(s) for quantification of transcription data, and especially for relative studies of the expression of target gene(s), is critical and should be carefully considered during the study design. The intuitive choice of GAPDH as a common reference gene caused a major shift in the quantitative expression data of target genes, inverting the relative expression values. According to the subtype of thyroid cancer and, of course, the target genes in this investigation, appropriate reference genes were proposed. An equation was developed based on this, and it was used to perform statistical analysis on over 25,000 genes. It is worth noting that while choosing a reference gene, the mean gene expression and its standard deviation should be carefully addressed. ![]() When eight common reference genes were examined, it was discovered that some of them not only lacked consistent expression but also had considerable expression variance. ![]() At the same time, meta-analyses were conducted in various ethnic groups, genders, and thyroid cancer subtypes. Using the relative RT-qPCR approach, we evaluated the expression of three target genes. Routine tissue-specific reference genes are often used in expression studies, but target genes are not taken into account.
0 Comments
Leave a Reply. |
AuthorWrite something about yourself. No need to be fancy, just an overview. ArchivesCategories |