Gelite™ Safe DNA Gel Stain 10,000X DMSO Solution

<strong>Comparison of DNA detection in 1% agarose gel in TBE buffer using Gelite™ Safe, EtBr, and SYBR® Safe.</strong> Two-fold serial dilutions of 1 kb DNA ladder were loaded in amounts of 100 ng, 50 ng, and 25 ng from left to right. Gels were stained for 60 minutes with Gelite™ Safe, EtBr, and SYBR® Safe according to the manufacturer's recommended concentrations and imaged using the ChemiDoc™ Imaging System (Bio-Rad®). Gels were illuminated using a 300 nm transilluminator fitted with a GelGreen filter.

Comparison of Gelite™ Safe (1:25,000X dilution) and GelRed® (1:10,000X dilution) in precast gel staining using 1% agarose gel in TBE buffer. Two-fold serial dilutions of 1 kb DNA ladder were loaded in the amounts of 100 ng, 50 ng, 25 ng, 12 ng, 6 ng, 3 ng, 1.5 ng, and 0.7 ng from left to right. Gels were imaged using a 300 nm transilluminator in ChemiDoc™ Imaging System (Bio-Rad®).

Stability test on Gelite™ Safe DNA Gel Stain, which was dissolved in DMSO and stored at four different temperatures (-20°C, 4°C, room temperature, and 37°C) for a period of one month. After storage, DNA ladders (100, 50, and 25 ng) were loaded in 1% agarose gel for 60 minutes at 75 V. Gels were incubated with Gelite™ Safe DNA Gel Stain for 1 hour. The image was captured in SYBR green filter set (Exposure time= 1.5 sec).

Comparison of DNA detection in 1% agarose gel in TBE buffer using Gelite™ Safe and GelRed®. Two-fold serial dilutions of 1 kb DNA ladder were loaded in amounts of 100 ng, 50 ng, and 25 ng from left to right. Gels were stained for 60 minutes with Gelite™ Safe and GelRed™ (Biotium) according to the manufacturer's recommended concentrations and imaged using the ChemiDoc™ Imaging System (Bio-Rad®). Gels were illuminated using a 300 nm transilluminator fitted with an EtBr filter set.

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Telephone	1-800-990-8053
Fax	1-800-609-2943
Email	sales@aatbio.com
Quotation	Request
International	See distributors
Shipping	Standard overnight for United States, inquire for international

Physical properties

Molecular weight	N/A
Solvent	Water

Spectral properties

Absorbance (nm)	509
Excitation (nm)	513
Emission (nm)	552

Storage, safety and handling

H-phrase	H303, H313, H333
Hazard symbol	XN
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R22
Storage	Room temperature (10-25 °C); Minimize light exposure
UNSPSC	12171501

Alternative formats

Gelite™ Safe DNA Gel Stain *10,000X Water Solution*

Gelite™ Safe DNA Gel Stain *GelRed Replacement, 10,000X in water*

Gelite™ Safe DNA Gel Stain *GelRed Replacement, 10,000X in DMSO*

Gelite™ Safe DNA Gel Stain *GelRed Alternative, 10,000X in water*

Gelite™ Safe DNA Gel Stain *GelRed Alternative, 10,000X in DMSO*

Overview SDS Protocol

Molecular weight

N/A

Absorbance (nm)

509

Excitation (nm)

513

Emission (nm)

552

AAT Bioquest is committed to designing our products to be environment-friendly. It is part of how we enable our customers to make the world healthier, cleaner, and safer. Ethidium bromide (EtBr) has been commonly used as a DNA stain for many years. However, EtBr is harmful if swallowed and is very toxic if inhaled. EtBr has been shown to be mutagenic in various tests and is an aquatic toxin. SYBR® Safe was introduced as a safer alternative to EtBr and SYBR® Green, but unfortunately, it is much less sensitive than SYBR® Green. It only has sensitivity comparable to EtBr. Gelite™ Safe has been developed specifically to be less hazardous than EtBr for staining DNA in agarose and acrylamide gels with much higher sensitivity. Gelite™ Safe has greatly improved safety and uncompromised sensitivity. The exceptional sensitivity and strong DNA binding affinity of Gelite™ Safe allows DNA to be stained prior to or post electrophoresis without destaining. In addition to its superior binding properties, Gelite™ Safe is essentially non-fluorescent in the absence of nucleic acids showing very low background fluorescence. Upon binding to nucleic acids, Gelite™ Safe exhibits a considerable fluorescence enhancement by several orders of magnitude greater than that of EtBr. Gelite™ Safe was optimized to be compatible with various instruments, including UV and blue-light transiluminators, gel documentation systems, and laser scanners. It is the first single formulation that can be used in either the green or red channel at your preference. Unlike the membrane-permeant SYBR® Green, which is highly toxic to cells and the environment, the membrane-impermeant properties of Gelite™ Safe make it a much safer and noncytotoxic alternative. Furthermore, Ames testing has confirmed Gelite™ Safe to be significantly less mutagenic than EtBr and SYBR® Green, even at concentrations well above the working concentration used for gel staining. Ames mutagenicity test was performed in a dose-dependent manner for all test dyes pretreated with an S9 fraction from rat liver (SYBR® is a trademark of ThermoFisher).

Platform

Gel Imager

Excitation	UV Transilluminator/Blue laser
Emission	SYBR® filter, GelStar® filter, GelGreen® filter, or GelRed® filter

Example protocol

PREPARATION OF WORKING SOLUTION

Gelite™ Safe working solution

Make 1X Gelite™ Safe working solution by diluting the 10,000X stock reagent with a buffer of your choice in a pH range of 7.5-8.5 (e.g., TAE, TBE or TE preferably pH 8.2).
Note Staining solutions prepared in water are less stable than those prepared in buffer and must be used within 24 hours to ensure maximal staining sensitivity.

SAMPLE EXPERIMENTAL PROTOCOL

The following protocols are recommended. However, some comparisons might be made to determine which one better meets your needs.

Post-staining protocol

Run gels according to your standard protocol.
Place the gel in a suitable polypropylene container. Gently add a sufficient amount of the 1X staining solution to submerge the gel.
Note Do not use a glass container, as it will adsorb much of the dye in the staining solution.
Agitate the gel gently at room temperature for ~30 to 60 minutes. Protect the staining container from light.
Note Destaining is not required. Image can be acquired without any wash steps.
Image the gel with a 300 nm/254 nm ultraviolet transilluminator, or a laser-based gel scanner using a long path green filter such as a SYBR® filter, GelStar® filter, GelGreen® filter, or GelRed® filter.

Pre-staining protocol

Prepare agarose gel solution using your standard protocol.
Dilute the 10,000X Gelite™ Safe stock reagent into the gel solution at 1:10,000 just prior to pouring the gel and mix thoroughly.
Run gels according to your standard protocol.
Image the gel with a 300 nm/254 nm ultraviolet transilluminator, or a laser-based gel scanner using a long path green filter such as a SYBR® filter, GelStar® filter, GelGreen® filter, or GelRed® filter.

Spectrum

Open in Advanced Spectrum Viewer

Spectral properties

Absorbance (nm)	509
Excitation (nm)	513
Emission (nm)	552

Images

Figure 1. Comparison of DNA detection in 1% agarose gel in TBE buffer using Gelite™ Safe, EtBr, and SYBR® Safe. Two-fold serial dilutions of 1 kb DNA ladder were loaded in amounts of 100 ng, 50 ng, and 25 ng from left to right. Gels were stained for 60 minutes with Gelite™ Safe, EtBr, and SYBR® Safe according to the manufacturer's recommended concentrations and imaged using the ChemiDoc™ Imaging System (Bio-Rad®). Gels were illuminated using a 300 nm transilluminator fitted with a GelGreen filter.

Figure 2. Comparison of Gelite™ Safe (1:25,000X dilution) and GelRed® (1:10,000X dilution) in precast gel staining using 1% agarose gel in TBE buffer. Two-fold serial dilutions of 1 kb DNA ladder were loaded in the amounts of 100 ng, 50 ng, 25 ng, 12 ng, 6 ng, 3 ng, 1.5 ng, and 0.7 ng from left to right. Gels were imaged using a 300 nm transilluminator in ChemiDoc™ Imaging System (Bio-Rad®).

Figure 3. Stability test on Gelite™ Safe DNA Gel Stain, which was dissolved in DMSO and stored at four different temperatures (-20°C, 4°C, room temperature, and 37°C) for a period of one month. After storage, DNA ladders (100, 50, and 25 ng) were loaded in 1% agarose gel for 60 minutes at 75 V. Gels were incubated with Gelite™ Safe DNA Gel Stain for 1 hour. The image was captured in SYBR green filter set (Exposure time= 1.5 sec).

Figure 4. Stability test on Gelite™ Safe DNA Gel Stain, which was dissolved in DMSO and stored at four different temperatures (-20°C, 4°C, room temperature, and 37°C) for a period of three months. After storage, DNA ladders (100, 50, and 25 ng) were loaded in 1% agarose gel for 60 minutes at 75 V. Gels were incubated with Gelite™ Safe DNA Gel Stain for 1 hour. The image was captured in SYBR green filter set (Exposure time= 1.5 sec).

Figure 5. Comparison of DNA detection in 1% agarose gel in TBE buffer using Gelite™ Safe and GelRed®. Two-fold serial dilutions of 1 kb DNA ladder were loaded in amounts of 100 ng, 50 ng, and 25 ng from left to right. Gels were stained for 60 minutes with Gelite™ Safe and GelRed™ (Biotium) according to the manufacturer's recommended concentrations and imaged using the ChemiDoc™ Imaging System (Bio-Rad®). Gels were illuminated using a 300 nm transilluminator fitted with an EtBr filter set.

Comparison of DNA detection in 1% agarose gel in TBE buffer using Gelite™ Safe, SYBR® Safe, and EtBr. Two-fold serial dilutions of 1 kb DNA ladder were loaded in amounts of 86 ng, 43 ng, 21.5 ng, 10.7 ng, 5.3 ng, 2.6 ng, 1.3 ng, and 0.5 ng from left to right. Gels were imaged using a 300 nm transilluminator in ChemiDoc™ Imaging System (Bio-Rad®).

Figure 6. Comparison of DNA detection in 1% agarose gel in TBE buffer using Gelite™ Safe, SYBR® Safe, and EtBr. Two-fold serial dilutions of 1 kb DNA ladder were loaded in amounts of 86 ng, 43 ng, 21.5 ng, 10.7 ng, 5.3 ng, 2.6 ng, 1.3 ng, and 0.5 ng from left to right. Gels were imaged using a 300 nm transilluminator in ChemiDoc™ Imaging System (Bio-Rad®).

$Summary of Ames test results. Ames mutagenicity test was performed in a dose-dependent manner for Gelite™ Safe, SYBR® Green, and EtBr. Samples were pretreated with an S9 fraction liver extract and then tested. With <em>S. Typhimurium</em> strain TA1538, an increase in revertants of more than two-fold over the background indicates a positive result for mutagenicity.$

Figure 7. Summary of Ames test results. Ames mutagenicity test was performed in a dose-dependent manner for Gelite™ Safe, SYBR® Green, and EtBr. Samples were pretreated with an S9 fraction liver extract and then tested. With S. Typhimurium strain TA1538, an increase in revertants of more than two-fold over the background indicates a positive result for mutagenicity.

Citations

View all 1 citations: Citation Explorer

BioAI for Anti-Infective Drug Discovery
Authors: Esquivel, Maria and Fernando, Johann and Fisher, Anna and Leong, Cameron and Weaver, Adam
Journal: (2022)

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