DESCRIPTION

TOROGreen DNA Staining Dye (10000×) is designed to be a safer replacement for conventional EtBr which poses a significant  health and safety hazard for its users. This dye is far more sensitive and stable than EB or SYBR series dye for staining dsDNA, ssDNA in agarose or polyacrylamide gels.This dye can be used to stain DNA in agarose gels via either precast or post gel staining, and in in polyacrylamide gels via post gel staining. TOROGreen DNA Staining Dye is compatible with a 470nm blue LED light transilluminator and imaging systems.

KIT CONTENTS and STORAGE

 500ul×5tubes . Store in the dark at room temperature.

FEATURES

•A safer alternative to EtBr because of nonmutagenic and noncytotoxic.

•Ultra-sensitive and extremely stable than other nucleic acid staining dye.

•Easy disposal: direct disposal down the drain or in regular trash.

•Compatible with a 470nm blue light transilluminator and imaging systems: increased cloning efficiency.

 PROTOCOL

Because high molecular weight and high affinity nucleic acid binding dyes can affect DNA migration during electrophoresis,post-staining of gels is highly recommended.Agarose gels can be precast with TOROGreen DNA Staining Dye ,however, it may affect the migration or resolution of some samples in precast gels. The precast protocol is not recommended for polyacrylamide gels.

Before opening, spin down the content of the tube to ensure the solution is at the bottom.

1. Post-Staining Gels

11.1 Perform agarose or polyacrylamide gel electrophoresis as usual according to your standard protocol.
1.2 Dilute TOROGreen DNA Staining Dye10000×stock solution 3300 fold to make a 3× staining solution in 0.1 M NaCl aqueous solution.
1.3 Carefully place the agarose or polyacrylamide gel in a suitable container such as a polypropylene staining tray. Add a sufficient amount of the 3× staining solution to submerge the agarose or polyacrylamide gel.
1.4 Agitate the agarose or polyacrylamide gel gently at room temperature for ~30 minutes.
     Note: Optimal staining time may vary somewhat depending on the thickness of the agarose gel and the percentage of agarose. For polyacrylamide gels containing 3.5-10% acrylamide, typical staining time is 30 minutes to 1 hour with gels of higher acrylamide content requiring longer staining time.
1.5 Destaining is not required, but the gel can be washed in water to reduce background if necessary.
1.6 View the stained gel with a 470nm blue LED light imaging systems or transilluminator.
1.7 Staining solution can be reused at least 2-3 times. Store staining solution at room temperature protected from light.

2.2.Precasting Agarose Gels
2.1 Prepare molten agarose gel solution using your standard protocol.
Note: the precast protocol is not recommended for polyacrylamide gels. Polyacrylamide gels can be stained using the post-stain protocol.
2.2 Dilute the TOROGreen DNA Staining Dye 10000× stock solution into the molten agarose gel solution at 1:10000 and mix thoroughly.The TOROGreen DNA Staining Dye can be added while the gel solution is still hot.
2.3 Cast the agarose gel and allow it to solidify. Load samples and perform agarose o gel electrophores is as usual according to your standard protocol.
2.4 View the stained gel with a 470nm blue LED light imaging systems or transilluminator.
2.5 Precast the gels containing TOROGreen DNA Staining Dye can be stored for future use for up to a month. We recommend storing gels at room temperature in the dark.Storage of precast gels at 4℃ can cause dye precipitation and poor performance.