Question

What could be causing a persistent interference in my gel analysis despite troubleshooting efforts, specifically on a Novex Wedgewell 6% Tris-Glycine Gel, where 10ul of Novex 2X Native Tris-Glycine Sample Buffer and 10ul of my sample were loaded in each well, and the gel was run at 100V for 1hr at 4C, as observed under a Fluorescein Blot filter?

Answer 1

Persistent interference in gel analysis could be due to contamination, degradation of reagents, or errors in gel staining and imaging protocols including the use of a Fluorescein Blot filter.

Step-by-step explanation:

Persistent interference in gel analysis using a Novex Wedgewell 6% Tris-Glycine Gel can arise from a variety of sources. Since multiple troubleshooting efforts have already been undertaken, it's possible that the issue could stem from faulty reagents or improper sample preparation.

It's critical to confirm the purity and quality of the Novex 2X Native Tris-Glycine Sample Buffer and your sample, as contamination can lead to unexpected results under a Fluorescein Blot filter. It's also important to check the gel itself for any inconsistencies in the polymerization or potential degradation.

Another common issue could be the detection method. For example, the usage of a Fluorescein Blot filter suggests that fluorescent labeling or staining is involved. Problems could arise from incomplete staining, inefficient washing steps, or a mismatched filter set leading to suboptimal fluorescence detection.