An approximate correlation between cartridge size, culture volume and the amount of DNA is given in the table below. Before starting with the isolation of Lambda DNA prepare buffer T2. Heating the solution may facilitate this process. The T2 buffer should be stored at 4C and used cold. Dissolve precipitated SDS by warming to 40C before use, if necessary.
All volumes given in the protocol are minimum volumes.
The yield of lambda DNA depends on several properties like E. coli strain, particular lambda vector, titer of the lambda lysate, efficiency of inoculation, and culture conditions. Transfer the supernatant to a clean tube. For the preparation of diluted phosphoric acid add 3-4 drops to 2 ml of water. Repeat this step if residual extraction matrix is visible. Discard the supernatant and drain the pellet by standing the tube in a tilted position for 3 min.