Abstract：
:Allelic differential expression (ADE) is common in diploid organisms, and is often the key reason for specific phenotype variations. Thus, ADE detection is important for identification of major genes and causal mutations. To date, sensitive and simple methods to detect ADE are still lacking. In this study, we have developed an accurate, simple, and sensitive method, named fluorescence primer PCR-RFLP quantitative method (fPCR-RFLP), for ADE analysis. This method involves two rounds of PCR amplification using a pair of primers, one of which is double-labeled with an overhang 6-FAM. The two alleles are then separated by RFLP and quantified by fluorescence density. fPCR-RFLP could precisely distinguish ADE cross a range of 1- to 32-fold differences. Using this method, we verified PLAG1 and KIT, two candidate genes related to growth rate and immune response traits of pigs, to be ADE both at different developmental stages and in different tissues. Our data demonstrates that fPCR-RFLP is an accurate and sensitive method for detecting ADE on both DNA and RNA level. Therefore, this powerful tool provides a way to analyze mutations that cause ADE.
journal_name
Sci Repjournal_title
Scientific reportsauthors
Zhao C,Xie S,Wu H,Luan Y,Hu S,Ni J,Lin R,Zhao S,Zhang D,Li Xdoi
10.1038/s41598-019-42815-5subject
Has Abstractpub_date
2019-04-19 00:00:00pages
6334issue
1issn
2045-2322pii
10.1038/s41598-019-42815-5journal_volume
9pub_type
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