Hi Neha Khandelwal , There are a few things to be clear before answering your question, why are you amplifying a gene from the genome? What is the main purpose? Is it for cloning for overexpression? When you are amplifying a gene from the gDNA of a higher animal it contains both exon and introns. For example, human IRF4 contains 9 exons, and 8 introns and the total length of the gene will be 19.4kbp. What is the purpose of cloning this? However, for the gDNA, the gene is there in the chromosome. It doesn’t matter the copy number for amplification. But, If you want to get functional CDS the story is different. There you need to isolate total RNA from cells, and then synthesize cDNA from it. The cDNA can use as a template to amplify the CDS of the gene. As you mentioned the copy number, you can use qPCR to determine the expression level of the gene of interest in different cell lines. At the same time you can do stimulation cells using some chemicals such as poly I: C. Recently, we have published a paper on the characterization of fish IRF 4, you can read it to get a clear idea (https://www.sciencedirect.com/science/article/pii/S0145305X22000180).

Hi Thilina, Thank you so much for your answer. yes, you are right. I have now ordered cDNA to amplify the CDS of the gene.