gDNA Amplification

[Mehmet Hazirci]

Hi All, 

I have recently extracted genomic DNA from all of my samples and I have got WAY more genomic DNA than the expected 6.6 pg/cell. In some samples, I have over 6 milligrams of DNA. The Nature protocols Joung et al 2017 paper says to amplify ALL of the genomic DNA extracted for each sample, however, given the number of samples I plan to sequence, and replciates, this would be over 20,000 PCR reactions (@ 7ug DNA/reaction) ad cost over £30,000 in NEBNext Mastermix alone. Is it 100% necessary to amplify all DNA from the sample. I have seen publications amplifying only 150 ug DNA per sample. I could even amplify 6.6pg x 500 x 123411 (500x coverage of each sgRNA) = 407 ug per sample.

Any help/advice appreciated. 

Best,

Mehmet