1.putting the 2µl cDNA on the wall
2.adding to solution
Which way is more efficient?
I usually added the template DNA into the master mix. So you can avoid variations in the template by pipetting errors. The downside of this is, that it costs a bit of cDNA (the overhead of the master mix).
Thank you christian.as u know for each marker I will have normal duplicates and tumor duplicates plus NTC! I make master mix fisrt and after distributing the last step will be adding template!
Add your cDNA for 2 tubes with nuclease free water and then divide it into 2 tubes.
@Nirmala: Thats the other possible way. I would go for a volume as big as possible (its easier to loose 0,5ul than 5ul).
I usually make master mix of buffer and primers of each marker in one tube and NFW and cDNA in another tube according to the number of wells you need to work with. Then you can aliquote as per your requirement.
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