Bluejuice gel loading buffer 10

The complexation efficiency was investigated using a 1% Agarose gel in Tris/Borate/EDTA buffer. Both running buffer and gel, contained SafeView (NBS Biologicals, Huntingdon, UK), a Nucleic Acid Stain (6 µL/100 mL). Free siRNA (positive control) and samples contained 3 µL BlueJuice^TM Gel Loading Buffer (10×) (Invitrogen, Waltham, MA, USA) and 27 µL sample. The gel was run at 90 V for 60 min (Bio-Rad, PowerPacBasic, Hercules, CA, USA) and imaged under UV with Uvitec Cambridge Mini HD.

To assess the behaviour of the NPs in the presence of serum, samples were exposed to 10% fetal bovine serum (FBS, heat-inactivated) for different time intervals at 37 °C for up to 24 h. Particle aggregation was monitored by DLS.
To assess the ability of the CD.NPs to prevent siRNA degradation following exposure to serum, a heparin displacement assay was used. Following incubation at 37 °C in 10% FBS, samples were treated with freshly prepared heparin (10 mg/mL, roughly 2000 I.U. in phosphate-buffered saline) for 75 min at room temperature (20% heparin in final sample). BlueJuice^TM Gel Loading Buffer (10×) (Invitrogen, Invitrogen, Waltham, MA, USA) was added to the samples and loaded onto the gel, containing SafeView (NBS Biologicals, Huntingdon, UK). The gel was run at 120 V for 30 min (Bio-Rad, PowerPacBasic, Hercules, CA, USA) and imaged under UV. The heparin-treated samples were measured with NanoDrop One (ThermoScientific, Dublin, Ireland) to quantify the residual siRNA.