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Table 1 presents the QPCR experiment steps

From: The variation in promoter sequences of the Akt3 gene between cow and buffalo revealed different responses against mastitis

No. Step process
1 Total RNA extraction.
2 Confirm the good purity and quantity of RNA, with 260/280 nm > 1.8 and >200 ng yield by NanoDrop spectrophotometer.
3 cDNA strand Synthation using cDNA kit
4 QPCR primer design using premier 5.0 for respective genes and confirmation of their accuracy through bioinformatics UCSC software.
5 The QPCR of the cDNA was performed by using master mix probe and the previously designed and checked primers. The protocol was as follows:
95 °C for 5 min, denaturing cycles as 94 °C for 30 s, annealing as 30-s temperature (Tm), extending at 30 s for 72 °C, with a final extension of 10 min at 72 °C.
6 Control group in experiment as after normalization of mRNA level and as a control group of actin.
7 The experiment samples were conducted in three replicates.