I usually forget to add proper amount of PCR ingredients and could not get proper results. I need the proper ingredients in proper quantity for 20 ul and 50ul PCR volume??
The basic ingredients for every PCR is the same. But, each ingredient's volume for setting up a reaction can be different. It depends on the concentration of your primers, the concentration of PCR buffer (ex. some with 5x and some with 10x).
See the protocol of setting up a reaction from using Phusion® High-Fidelity DNA polymerase (https://www.neb.com/protocols/1/01/01/pcr-protocol-m0530). Usually, on the datasheet which comes with the DNA polymerase kit will have reaction preparation info.
Rather than focusing on volume, be sure that you know final oncentration of ingredients which you require in a PCR reaction. Whatever volume it is, or whatever is the concentration of atock solutions (it may vary from lab to lab), just use formula C1V1=C2V2. So if you know final concentrations of primers, dNTPs, buffer, MgCl2, template, Taq in reaction, use the formula and here you go.
By the way, you said 'you want a proper ingredients'.
Except the basic ingredients for PCR, some people prefer to add some 'PCR additives' (ex. DMSO, which enhances PCR specificity), some people don't need it (because the PCR buffer they purchase has already formulated with it). So, it also depneds on the DNA template (GC-rich vs. non GC-rich) you want to amplify and which type of kit you use to prepare the reaction ingredients.
As every one else have pointed out you need to focus on final concentration of all the ingredients in the PCR recipe.Typically it comprise of
10X KCl complete PCR buffer (with MgCl2) 1.2 µl (1X)
10mM dNTP 0.25 µl (200μM)
0.1%BSA 1.2 µl (0.01%)
Taq 0.05ul(1U)
D/W 5.2 μl
Specific primers 10-to 50 pM
Total volume per reaction 10.0 μl
To this add 2ul of your templet. Final volume 12ul
Hope this helps
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