IGEM:Harvard/2006/Gel staining

EtBr staining of mini PA gels

Incubate in 10μL EtBr + 100 mL Tris-glycine buffer for 30 min.

Coomassie staining of mini PA gels

Protocol from Pierce using GelCode Blue:

• Wash
  • Native PAGE: Rinse gel with ultrapure water for 5 minutes.
  • Note: Gels electrophoresed with MOPS or MES running buffers must be prefixed with a 50% methanol and 7% acetic acid solution for 15 minutes and then washed with ultrapure water to remove fixing solution. GelCode ® Blue Stain penetrates prefixed gels better than non-fixed gels and, therefore, standard SDS-PAGE gels may also be prefixed with good results.

• Stain Note: Mix the GelCode ® Blue Stain Reagent solution immediately before use by gently inverting or tipping and swirling the bottle several times. Such mixing is especially important when using Product No. 24592 with a dispenser pump. Do not shake bottle to mix the solution.

• • Add 20 ml of GelCode ® Blue Stain Reagent for an 8 x 10 cm mini gel. Additional reagent may be required if a large tray is being used. Gently shake tray and periodically monitor protein band development. Stain intensity reaches a maximum within approximately 1 hour. Gels may be stained overnight without increasing background.

• • Note: PhastGel ® Gels may require increased staining times (2 hours to overnight) for optimal band development.

• Destain (Water Wash EnhancementTM Step) Replace Stain Reagent with ultrapure water. Several water changes for a 1-2 hour period may be necessary for optimal results. This step enhances stain sensitivity, as weak protein bands continue to develop.