SAFELOOK™
This product was developed as a nucleic acid reagent to replace ethidium bromide, which is known to be highly mutagenic. SAFELOOK™ Green/Red Nucleic Acid Stain can be used both pre- and post-staining. SAFELOOK™ Load-Green/Load-Red is mixed with samples at the time of use.
Features
- Safe to use because of its low mutagenicity
- Two types of products are available: pre-/post-staining and loading-dye
Product selection chert
SAFELOOK™ Green Nucleic Acid Stain |
SAFELOOK™ Red Nucleic Acid Stain |
SAFELOOK™ Load-Green (6×) |
SAFELOOK™ Load-Red (6×) |
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Staining method | Pre-/Post-staining | Pre-/Post-staining | Addition to samples | Addition to samples |
---|---|---|---|---|
Excitation wavelength | 490 nm | 540 nm | 490 nm | 540 nm |
Fluorescence wavelength | 520 nm(DNA) 635 nm(RNA) |
630 nm | 525 nm | 630 nm |
Light source | LED*1 / UV | LED / UV*2 | LED*1 / UV | LED / UV*2 |
Recommended gel | Agarose | Agarose | Agarose/Polyacrylamide | Agarose/Polyacrylamide |
Recommended sample | dsDNA / ssDNA / RNA | dsDNA / ssDNA / RNA | dsDNA / ssDNA / RNA | dsDNA / ssDNA / RNA |
- 1: SAFELOOK™ Green Nucleic Acid Stain and SAFELOOK™ Load-Green (6x) can be observed under illumination from a blue LED.
- 2: We recommend that SAFELOOK™ Red Nucleic Acid Stain and SAFELOOK™ Load-Red (6x) are observed under UV. (If you choose observation under LED illumination, please ensure suitable conditions)
Amount to use
Pre-staining
Add 5 µL of SAFELOOK™ per 100 mL of gel solution. As required, add a further 5-10 µL to 200 mL of electrophoresis buffer.
Post-staining
Add 10-20 µL of SAFELOOK™ per 100 mL of buffer (1:5000-1:10000).
Addition to samples
Add SAFELOOK™ to samples/markers in the ratio of 1:5.
Examples of Use
SAFELOOK™ Green Nucleic Acid Stain |
SAFELOOK™ Red Nucleic Acid Stain |
SAFELOOK™ Load-Green (6×) |
SAFELOOK™ Load-Red (6×) |
|||||
Pre-staining | Post-staining | Pre-staining | Post-staining | Addition to samples | Addition to samples | |||
LED | UV | LED | UV | UV | UV | LED | UV | UV |
Staining methods
- Pre-staining: During gel preparation, 5 μL of staining reagent was added per 100 mL of agarose gel solution. 5 μL of staining reagent was also added per 100 mL of electrophoresis buffer.
- Post-staining: 10 μL of staining reagent was added per 100 mL of TAE buffer, to prepare the staining buffer. After electrophoresis, gels were soaked in the staining buffer for 30 minutes.
- Addition to samples: Staining reagent and samples were mixed in the ratio of 1:5. The mixture was applied to agarose gel, and electrophoresis was performed.
Comparison with ethidium bromide (EtBr)
Agarose electrophoresis of a DNA marker (Product Number: 313-06961) was performed under the same conditions; bands were observed using an LED or a UV transilluminator. Staining was performed in accordance with the conditions recommended for each staining reagent (SAFELOOK™ or EtBr).
SAFELOOK™ Green Nucleic Acid Stain | SAFELOOK™ Red Nucleic Acid Stain | ||||||
Pre-staining/LED | Post-staining/LED | Pre-staining/UV | Post-staining/UV | ||||
SAFELOOK™ | EtBr | SAFELOOK™ | EtBr | SAFELOOK™ | EtBr | SAFELOOK™ | EtBr |
Q&A
Q. Mobility of bands is altered.
A.With pre-staining, the dye binds to the nucleic acid and reduces mobility. Especially when samples contain large amounts of nucleic acid, or when there is a difference in the concentration of nucleic acid among lanes, mobility may differ even among nucleic acids of the same molecular weight. With pre-staining, the dye binds to the nucleic acid and reduces mobility. Especially when samples contain large amounts of nucleic acid, or when there is a difference in the concentration of nucleic acid among lanes, mobility may differ even among nucleic acids of the same molecular weight.
One effective solution is to change the staining method to post-staining; however, the problem may be solved simply by diluting the sample. The salt concentration of the sample also affects band mobility. In such circumstances, ethanol precipitation or re-preparation of the sample using a desalting column may solve the problem.
Q. How can I lower the background intensity?
A. The background intensity can be lowered by destaining the gel for a short time in water, before putting the gel into a gel observation system. However, destaining is usually not necessary.
Q.Can blue light (LED) also be used for observation?
A.SAFELOOK™ Green Nucleic Acid Stain and SAFELOOK™ Load-Green (6x) can be observed under blue light illumination.
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Pre/Post-staining Type
Loading Dye Type
For research use or further manufacturing use only. Not for use in diagnostic procedures.
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