I agree with Paul. You must decide what kinf of qPCR will you do, quantification or relative? If you want to compare the gene expression of one sample with another, you need only positive controls, just to be sure about the qPCR protocol. If you want to quantify your sample, you'll need a positive control, however with known concentration. In both cases, the control is the sample that works for these primer pairs. The standard is used to produce the standard curve, so to be able to quantify your samples. For optimisation of the primers, you need just a sample, in which the gene you study is expressed.

Hi Rukshana

Standards are what you will use to quantify your target genes. These are generally DNA or cloned target gene of a known concentration is diluted over a few orders of magnitude. You can then use these to determine the concentration of your unknown sample.

Controls are the same as for any PCR. a negative and a positive to validate that your products are genuine.

Are you using Sybr for the qPCR experiment. I would be happy to help you some more with the experimental details.

I agree with Paul. You must decide what kinf of qPCR will you do, quantification or relative? If you want to compare the gene expression of one sample with another, you need only positive controls, just to be sure about the qPCR protocol. If you want to quantify your sample, you'll need a positive control, however with known concentration. In both cases, the control is the sample that works for these primer pairs. The standard is used to produce the standard curve, so to be able to quantify your samples. For optimisation of the primers, you need just a sample, in which the gene you study is expressed.

Thanks a lot! I will be using SYBR Green. And its for checking expression levels, yes. so, I guess, I need to use just the positive and negative controls?