Home FAQs After incubation with NEBExpress® T4 Lysozyme, the lysate became viscous; how can I resolve this?

FAQ: After incubation with NEBExpress® T4 Lysozyme, the lysate became viscous; how can I resolve this?

A viscous lysate results from an abundance of DNA released. To reduce viscosity, three options are available:

  1. Shear the DNA by subjecting the lysate to a couple of pulses on a sonicator.
  2. Add 1mM CaCl2 and 1 µL of Micrococcal Nuclease (NEB #M0247S) per 1 mL of lysate, mix, and incubate for 5 minutes at room temperature. 
  3. Add 1 µL of DNase I (NEB #M0303S) per 1 mL of lysate and 1 mM CaCl2 or 1 mM MgCl2 (especially if the suspension contains EDTA), mix, and incubate for 5 minutes at room temperature. NEB’s DNase I is tagged with maltose-binding protein (MBP), making its removal via amylose resin (NEB #E8021), amylose resin high flow (NEB #E8022), or amylose magnetic beads (NEB #E8035) simple and effective.