SafeRed^® Nucleic Acid Gel Stain (10000× in water)        

SafeRed^®, SafeView^®, SafeStain^®和GelStain^®,and DuGreen^®are registered trademarks and strictly forbidden to be used without authorization. 

Product Description

SafeRed^® (MW>1000) Nucleic Acid Gel Stain is our third generation nonmutagenic fluorescent nucleic acid gel stain to replace the highly toxic traditional ethidium bromide (EtBr) for staining dsDNA, ssDNA or RNA in agarose gels or polyacrylamide gels. The third part’s Ames test by WuXi Pharmacy shows that SafeRed is nonmutagenic for gel staining. DuGreen and SafeRed Nucleic Acid Gel Stain are highly sensitive than EtBr either as precast gel stains or post gel stains.

We can directly replace EB with SafeRed without changing the existing imaging system since SafeRed and EB have the similar spectra. SafeRed can also be used to stain dsDNA, ssDNA or RNA in polyacrylamide gel via post gel staining. Gel staining with SafeRed is compatible with downstream applications such as gel extraction and cloning. SafeRed is efficiently removed from DNA by phenol/chloroform extraction and ethanol precipitation.

Features

1) Relative Safety: Nonmutagenic and noncytotoxic

2) Easy disposal: Safe to dispose in the drain

3) Good Compatibility: Spectrally compatible with all the existing instruments

4) High Sensitivity: Higher signal but lower background

6) High Stability: can be stored at RT and microwavable

Precast Protocol for Agarose Gel Electrophoresis

Pouring a Standard 1% Agarose Gel:

1   Pour 1g of Agarose powder and 100mL of 1xTAE or 1xTBE into a glass flask.

Note:Agarose gels are commonly used in concentrations of 0.7% to 2% depending on the size of DNA bands.

2   Melt the agarose in a microwave for 1-3min (until the agarose is completely dissolved and the solution becomes clear, the solution is completely clear and no small floating particles are visible.)

Note:Caution HOT! Be careful!  Do not let the solution boil for long periods as it may boil out of the flask.

3   Let agarose solution cool down to about 50-55°C, Add 10uL SafeRed per 100mL gel, gently stirring the gel solution to mix thoroughly.

4   Seal horizontal gel apparatus. Pour molten agarose onto gel plate to a depth of 4~8 mm. Insert a comb until its base is 1 mm from the base of the gel. Allow cooling.

Note:Pour slowly to avoid bubbles, which will disrupt the gel. Any bubbles can be pushed away from the well comb or towards the sides/edges of the gel with a pipette tip.

5   Place newly poured gel at 4°C for 10-15 minutes OR let sit at room temperature for 20-30 minutes, until it has completely solidified. Remove comb and submerge in 4X TAE buffer 

Loading Samples and Running an Agarose Gel:

1   Once solidified, place the agarose gel into the gel box.

2   Add 1/5 volume 6X Loading buffer to sample. 

Note:Loading buffer serves two purposes: 1) it provides a visible dye that helps with gel loading and will also allows you to gauge how far the gel has run while you are running your gel; and 2) it contains a high % glycerol, so after adding it your sample is heavier than water and will settle to the bottom of the gel well, instead of diffusing in the buffer.

3   Fill gel box with 1xTAE (or TBE) until the gel is covered.

4   Carefully Pipette 5 ml of the DNA ladder standard into the first lane of the gel.

Note:When loading the sample in the well, maintain positive pressure on the sample to prevent bubbles or buffer from entering the tip. Place the very top of the tip of the pipette into the buffer just above the well. Very slowly and steadily, push the sample out and watch as the sample fills the well. After all samples are unloaded, push the pipettor to the second stop and carefully raising the pipette straight out of the buffer.

5   Carefully pipette 5 ml of each sample/Sample Loading Buffer mixture into other separate wells in the gel.

6   Run the gel at 80-150V untilBromophenol Blue is near the end of the gel. This dye runs at ~800b.

Note:Black is negative, red is positive. (The DNA is negatively charged and will run towards the positive electrode.) Note:A typical run time is about 0.5-1 hour, depending on the gel concentration and voltage.

7   Turn OFF power, Disconnect the electrodes from the power source, and then carefully remove the gel from the gel box.

8   Visualize or Imagethe stained gel with a standard transilluminator (302 or 312nm), and photograph the gel using an ethidium bromide filter. 

Post-Staining Protocol

1.       RunAgarose Gel Electrophoresisaccording to hereinbefore. 

2.       Dilute 10000X SafeRed stock solution to 1X staining solution in H₂O with 0.1M Nacl(e.g, add 5ul of SafeRed^®10,000X stock reagent and 5ml 1M Nacl to 45ml H₂O). Note: including 0.1M NaCl in the staining solution enhances sensitivity, but may promote dye precipitation if the gel stain is reused. (This solution can be reused at least 2~3 times protected from light).

3.       Using gloves, carefullyremove the gel from the casting tray and place into the staining dish, add 1X SafeRed staining solution to submerge the gel.

4.       Agitate the gel gently at room temperature for~30 minutesand the amount of the stain may depend on the thickness of the gel and the percentage of the agarose with gently shaking.

5.       Rinsing the gel with water can reduce the background. 

6.       Visualize or Imagethe stained gel with a standard transilluminator (302 or 312nm), and photograph the gel using an ethidium bromide filter. 

NOTE: Always wear lab coats, gloves and goggles when working with our products although they are low-risk chemicals for R&D only.