Can I amplify from plasma or serum samples using the REPLI-g Mini/Midi kits?
Plasma and serum will work, but these samples typically contain small numbers of cells, and small amounts of free DNA. Therefore, sufficient sample volume needs to be added to get enough cells to amplify. For our REPLI-g Whole Genome Amplification Services, we need about 50 ul of this sample type for successful amplification.
We suggest to concentrate plasma and serum samples to a final volume of 100-200 ul using centrifugal micro-concentrators. The Millipore 5 micron filter Ultrafree-CL SV will work fine for this purpose. Use 2.5 ul of the concentrate in the REPLI-g Mini/Midi kit reaction. If you use a minimum of 200-300 ul of sample for spin column concentration, you can remove 2.5 ul of the retained 10 ul for amplification using the kit. Please see the article Retention of leukocytes in capillaries: role of cell size and deformability by G. P. Downey et al. in the Journal of Applied Physiology 69(5), p. 1767-1778. It demonstrates that white cells will be retained on a 5 micron filter while red cells and platelets will filter through.
You can also try our Supplementary protocol “Whole genome amplification from plasma and serum using the REPLI-g® Midi Kit”