Hi everyone. I have performed a qPCR of OCT4, BMP4, ABCG2, CK3 gene and GAPDH as a reference gene. After I run all my samples, I just realized that NTC for OCT4 and CK3 showing Cq value but not the rest. Then I perform a qPCR for NTC only to all my primers. Only OCT4 and CK3 that showed Cq values. I was using primer design from previous student in my lab.
I contacted the supplier, they said it can be a contamination or a dimer. I believe is not the contamination. because I order two box, and i run the qPCR for NTC only for those 2 box, and the results is consistent. only OCT4 that showed a Cq value.
My question is can I still use the qPCR results of the OCT4?the data seems okay. some samples show high relative expression, some lower. Some showed higher Cq values compared to the NTC.
Using such data for the study is not advisable. In such scenario, working stock can be prepare again from original primer stock in hood and if you are still facing the same issue, you need to re order a new vial. Also use fresh plasticware and avoid using same pipettes (Used previously)while reconstitute the stock.
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