Nucleic Acid Stains Product Specification

5-Aminoallyl-dUTP can be enzymatically incorporated into DNA. The resulting amine-containing DNA can be subsequently labeled with a fluorescent dye, biotin or other haptens via conventional peptide coupling method.^1,2 This two-step method for labeling nucleic acids is considerably more economical than the one-step method using a prelabeled dUTP.³

• White solid soluble in water. Store at -20°C.
• C₁₂H₁₇N₃Na₃O₁₄P₃.
• Mwt: 589.17

AA-dUTP in lyophilized powder form is suitable for long-term storage.

5-Aminoallyl-dUTP can be enzymatically incorporated into DNA. The resulting amine-containing DNA can be subsequently labeled with a fluorescent dye, biotin or other haptens via conventional peptide coupling method.^1,2 This two-step method for labeling nucleic acids is considerably more economical than the one-step method using a prelabeled dUTP.³

• White solid soluble in water.Store at -20°C.
• C₁₂H₁₇N₃Na₃O₁₄P₃.
• Mwt: 589.17

5-Aminoallyl-UTP can be enzymatically incorporated into DNA. The resulting amine-containing DNA can be subsequently labeled with a fluorescent dye, biotin or other haptens via conventional peptide coupling method.

Colorless solution

Store at -20°C.

C₁₂H₁₇N₃Na₃O₁₅P₃.

Mwt: 605.17.

AA-UTP in lyophilized powder form is suitable for long-term storage. For more information see product information for 40021.

BrdU can be incorporated into DNA during cell division and subsequently detected by a BrdU antibody. The probe can be used to study cell-cycle kinetics.¹

White solid soluble in DMSO.

Store at -20°C and protect from ligh.

C₉H₁₁BrN₂O₅.

Mwt. = 307.10.

[59-14-3]

5-Bromo-dUTP is widely used in TUNEL assay to detect apoptosis cells,^1,2 and it is also a good substrate for reverse transcriptase.^3,4

Colorless solution

Store at -20°C and protect from light.

C₉H₁₁BrN₂Na₃O₁₄P₃.

Mwt. = 612.99.

BrUTP can be enzymatically incorporated into RNA.

Colorless solution

Store at -20°C and protect from light.

C₉H₁₁BrN₂Na₃O₁₅P₃.

Mwt. = 628.98.

7-AAD is a fluorescent DNA intercalator, which upon binding to dsDNA can be excited by the argon-ion laser and emits at 647 nm.^1-3 7-AAD binds selectively to GC regions of DNA, which makes the dye useful for chromosome banding studies.⁴

• l_ex\l_em(DNA) = 546/647 nm
• Orange red solid soluble in DMF or DMSO
• Store at -20°C and protect from light, especially in solution
• C₆₂H₈₇N₁₃O₁₆
• Mwt: 1270.45

Acridine orange (AO) stains dsDNA green (525 nm) and RNA or single stranded DNA red (650 nm).¹ The dye is membrane-permeant and its nucleic acid binding property has been used for cell-cycle studies.^2,3 Acridine orange has also been used for the detction of microorganisms in cerebrospinal fluid and other clinical specimens.⁴ We offer a highly purified form of acridine orange while most of the other commercially available grades of AO are either in zinc chloride complex form or of low purity.

l_ex\l_em= 500/526 nm

Yellow solution

Store at -20°C and protect from light

C₁₇H₂₀ClN₃

Mwt: 301.82

Actinomycin D is a nonfluorescent GC-selective intercalator.¹ Similar to 7-AAD, actinomycin D has also been used for chromosome banding studies.²

l_ex\l_em= 442nm/none

Orange red solid soluble in DMF or DMSO

Store at -20°C and protect from light, especially in solution

C₆₂H₈₆N₁₂O₁₆

Mwt: 1255.43

We developed aminooxy-biotin as an alternative to ARP (90073),^1-3 which is used for labeling abasic sites in damaged DNA. Aminooxy-biotin and ARP have similar reactivity toward aldehydes or ketones. However, aminooxy-biotin is hydrolytically more stable than ARP, and the longer spacer group in aminooxy-biotin should facilitate the interaction between biotin and avidin or streptavidin. Figure 7.11 shows the mechanism of the reaction.

• White solid soluble in DMSO
• Store at -20°C
• C₁₉H₃₂F₃N₅O₆S
• Mwt: 515.55

ARP reacts with the exposed aldehyde group formed at abasic sites in damaged DNA, allowing the DNA to be labeled with biotin groups.^1,2 The labeled DNA can then be quantitated with fluorescent or enzyme-conjugated streptavidin complexes. ARP can freely cross the cell membranes, thus allowing detection of abasic sites in living cells.³ ARP is suitable for use in microplate assays.

• White solid soluble in DMSO.
• Store at -20°C.
• C₁₄H₂₂F₃N₅O₆S.
• Mwt: 445.41.

Biotin-11-CTP can be enzymatically incorporated into RNA

• Colorless solution
• Store at -20°C and protect from light
• C₂₈H₄₂N₇O₁₇P₃SLi₄
• Mwt: 901.42

Biotin-11-dCTP can be enzymatically incorporated into DNA via nick translation, random priming, or 3'-end terminal labeling. The number ?11? is the number of atoms in the linker between biotin and dCTP.

• Colorless solution
• Store at -20°C and protect from light
• C28H42N7O16P3SLi4
• Mwt: 885.42

Biotin-11-dUTP can be enzymatically incorporated into DNA via nick translation, random priming, or 3'-end terminal labeling. The number '11' is the number of atoms in the linker between biotin and dUTP. Biotium also offers biotin-16-dUTP (40022 and 40022-1) and biotin-20-dUTP (40030 and 40030-1). The length of the linker affects the incorporation efficiency of the biotin-dUTP probe into DNA using DNA polymerases, and it also affects biotin/avidin or biotin/streptavidin. In general, the shorter the linker, the more efficiently the biotin-dUTP is incorporated into DNA by DNA polymerases. On the other hand, the longer the linker, the better biotin can interact with avidin or streptavidin.

Colorless solution

Store at -20°C and protect from light

C₂₈H₄₁N₆O₁₇P₃Sli₄

MWt: 886.5

This product is essentially the same as 40029 except that it is is a lyophilized solid that is more suitable for long term storage. The product contains lyophilized TE buffer, so you only need to add an appropriate amount of deionized H₂O to reconstitute the solution. For more product information, see 40029.

White solid soluble in H₂O

Store at -20°C and protect from light

C₂₈H₄₁N₆O₁₇P₃Sli₄

MWt: 886.5

Biotin-11-UTP can be enzymatically incorporated into RNA.

Colorless solution

Store at -20°C and protect from light

C₂₈H₄₁N₆O₁₈P₃SLi₄

Mwt: 902.5

Biotin-16-dUTP can be enzymatically incorporated into DNA via nick translation, random priming, or 3′-end terminal labeling.¹ The terminal deoxynucleotidyl transferase (TDT)-mediated biotin-dUTP nick end-labeling (TUNEL) method has been commonly used for apoptosis studies.^3,4 The number '16' is the number of atoms in the linker between biotin and dUTP. Biotium also offers biotin-11-dUTP (#40029 and #40029-1) and biotin-20-dUTP (#40030 and #40030-1). The length of the linker affects the incorporation efficiency of the biotin-dUTP probe into DNA using DNA polymerases, and it also affects biotin/avidin or biotin/streptavidin. In general, the shorter the linker, the more efficiently the biotin-dUTP is incorporated into DNA by DNA polymerases. On the other hand, the longer the linker, the better biotin can interact with avidin or streptavidin.

• Colorless solution
• Store at -20°C and protect from light
• C₃₂H₄₈N₇O₁₈P₃Sli₄
• MWt: 971.5

This product is essentially the same as 40022 except that it is is a lyophilized solid that is more suitable for long term storage. The product contains lyophilized TE buffer, so you only need to add an appropriate amount of deionized H₂O to reconstitute the solution. For more product information, see 40022.

• White solid soluble in H₂O
• Store at -20°C and protect from light
• C₃₂H₄₈N₇O₁₈P₃Sli₄
• Mwt: 971.5

Biotin-16-UTP can be enzymatically incorporated into RNA.

• Colorless solution
• Store at -20°C and protect from light
• C₃₂H₄₈N₇O₁₉P₃Sli₄
• MWt: 987.51

This product is similar to biotin-11-dUTP (40029) and biotin-16-dUTP (40022) except that it has a longer and water-soluble spacer group, which should facilitate interaction between the biotin group and avidin or streptavidin.

• Colorless solution
• Store at -20°C and protect from light
• C₃₃H₅₁N₆O₂₁P₃SLi₄
• Mwt: 1020.54

This product is essentially the same as 40030 except that it is is a lyophilized solid that is more suitable for long term storage. The product contains lyophilized TE buffer, so you only need to add an appropriate amount of deionized H₂O to reconstitute the solution. For more product information, see 40030.

• White solid soluble in H₂O
• Store at -20°C and protect from light
• C₃₃H₅₁N₆O₂₁P₃SLi₄
• Mwt: 1020.54

This product is similar to biotin-11-UTP (40033 ) and biotin-16-UTP (40023 ) except that it has a longer and watersoluble spacer group, which should facilitate interaction between the biotin group and avidin or streptavidin.

Colorless solution

Store at -20°C and protect from light

C₃₃H₅₁N₆O₂₂P₃SLi₄

Mwt: 1036.26

Because of the potential toxicity of DAPI, we offer DAPI dilactate dissolved in H₂O as a safer alternative to the powdery form, which is more likely for one to be in contact with. DAPI is a popular blue fluorescent DNA probe. The dye binds to the minor groove of dsDNA with a ~20-fold fluorescence enhancement. See Figure 9.3 for spectra.

l_ex\l_em(DNA) = 358/461 nm

Yellow solution

Store at -20°C and protect from light

C22H27N5O6

Mwt: 457.49

DAPI is a popular blue fluorescent DNA probe. The dye binds to the minor groove of dsDNA with a ~20-fold fluorescence enhancement. See Figure 9.3 for spectra.

• l_ex\l_em = 358/461 nm (DNA).
• Off-white solid soluble in H₂O.
• Store at 4°C and protect from light, especially in solution.
• C₁₆H₁₇Cl₂N₅.
• Mwt. = 350.25.
• [28718-90-3]

DAPI dilactate is essentially the same as DAPI (dihydrochloride) (40011 ) except for the difference in their counter ions. The dilactate counter ions make the dye more water soluble and therefore DAPI dilactate is a better choice if one wants to make the stock solution in deionized water. See Figure 9.3 for spectra.

• l_ex\l_em(DNA) = 358/461 nm
• Yellow solid soluble in water
• Store at 4°C and protect from light, especially in solution
• C₂₂H₂₇N₅O₆
• Mwt: 457.49

Dihydroethidium is the chemically reduced form of the commonly used DNA dye ethidium bromide. The probe is useful to detect oxidative activities in viable cells, including respiratory burst in phagocytes. Dihydroethidium itself is blue fluorescent (l_ex\l_em: 355/420nm) in cell cytoplasm while the oxidized form ethidium is red fluorescent (l_ex\l_em: 518/605 nm) upon DNA intercalation

• Off-white to light brown solid soluble in DMF or DMSO
• Store at -20°C. Under nitrogen or argon, and protect from light, especially when in solution.
• C₂₁H₂₁N₃
• Mwt: 315.
• [38483-26-0]

DiSC₂(5) binds to DNA-DNA or PNA-DNA duplexes with a large blue-shift of the absorption spectrum. Binding of the dye to PNA-DNA hybrid, in particular, blue-shifts the dye absorption spectrum by as much as 115 nm, offering a visual method to detect nucleic acid hybridization.^1,2

l_abs (pH7.5 buffer) = 646 nm (free dye); ~586 nm (dsDNA); ~531 nm (PNA-DNA)

Dark solid soluble in DMSO or MeOH

Store at 4°C

C₂₃H₂₃IN₂S₂

Mwt: 518.48

DODC has been reported to bind to triplex DNA as well as dimeric hairpin quadruplex DNA.^1,2 Binding of the dye to triplex DNA quenches the fluorescence of the dye and red-shifts the dye absorption peak.

l_abs = 582 nm

Red solid soluble in DMSO or MeOH

Store at 4°C

C₂₃H₂₃IN₂O₂

Mwt: 486.35

Because of its toxicity as a potent mutagen, we offer EB in H₂O as a safer alternative to the more hazardous powdery form, which is more likely for one to be in contact with. EB is an intercalating DNA-binding dye with little sequence preference. Once bound to nucleic acids, the fluorescence of the dye is enhanced by more than 10 times while the excitation maximum is redshifted by 30-40 nm and emission is blue-shifted by ~15 nm. EB is widely used as a nucleic acid gel stain.

l_ex\l_em(DNA) = 518/605 nm

Red solution

Store at 4°C and protect from light

C₂₁H₂₀BrN₃

Mwt: 394.31

Ethidium Homodimer, first developed by Dr. Le Pecq and his colleagues, is a high affinity fluorescent nucleic acid stain. It binds to both DNA and RNA in a sequence-independent manner and with a >30-fold fluorescence enhancement. The DNA binding of each Ethidium Homodimer covers four base pairs and is believed to occur by intercalation. Because the dye is highly positively charged, it can not cross cell membranes to stain living cells. However, it is very useful to detect nucleic acids in solution, or cells with disintegrated cell membranes. Biotium offers a high purity grade Ethidium Homodimer that is not available from anywhere else. Ethidium Homodimer from other suppliers often contains a high amount (as much as 20%) of inorganic salt that lowers the weight percent purity. 

• l_ex/l_em (with DNA) = 528/617 nm.
• l_ex(in H₂O, no DNA) = 493 nm. 
• Red solid soluble in DMSO or MeOH.
• Store at 4°C
• C₄₆H₅₀Cl₄N₈.
• Mwt: 857. 
• [61926-22-5]

Ethidium Homodimer supplied in a ready-to-use form. See 40010 for molecular information.

Ethidium Homodimer III was developed by Biotium as an alternative to Ethidium Homodimer I. It has absorption and emission spectra similar to those of Ethidium Homodimer I. However, the dye stains DNA 70% brighter than Ethidium Homodimer I. The dye is also a component of our Cell Viability/Cytotoxicity Assay Kit for detecting both live and dead cells in the same population (30002). See Figure 9.5 for spectra.

l_ex\l_em (with DNA) = ~530/~620 nm

Red solid soluble in DMSO, MeOH, or H₂O

Store at 4°C

Mwt:~1000

EthD-III in ready to use form. See 40050 for more information.

Ethidium monoazide bromide is a fluorescent nucleic acid stain with a photoaffinity label. The dye, after photolysis, binds covalently to nucleic acids.¹ The dye has been used to ?footprint? drug binding sites on DNA2 to modify plasmid DNA,^3,4 and to determine hemopoietic cell phenotype, function and position in the cell cycle.⁵ A particularly useful application of the dye is to selectively and covalently label dead cells in the presence of live cells. Since ethidium monoazide bromide is relatively impermeant to live cells, it selectively labels DNA in dead cells in a mixed population of live and dead cells. Photolysis following the dye application renders the dead cell DNA covalently labeled with the dye. One can then wash and fix the cell preparation and exam it by microscopy fluorescence plate reader or flow cytometry. The major advantage of this method is that researchers can avoid extensive manipulation of live pathogenic organisms.⁶

l_ex (pH3) = 458 nm7

Orange solid soluble in DMF, or ethanol

Store at -20°C and protect from light, especially in solution

C₂₁H₁₈BrN₅.

Mwt: 420

[58880-05-0]

Biotium in collaboration with AlleLogic Biosciences Corp. Has developed EvaGreen^TM as a superior fluorescent DNA stain for quantitative real-time PCR (qPCR). EvaGreen^TM is truly a remarkable dye in many aspects. Upon binding to DNA, the fluorescence of EvaGreen^TM is several-fold higher than that of SYBR Green I while the dye shows very little inhibition to the PCR process. Unlike SYBR Green, which has been reported to be unstable, EvaGreen^TM is highly robust, both thermally and hydrolytically under alkaline or acidic condition. In addition, the absorption and emission spectra of EvaGreen^TM are similar to those of SYBR Green I or FAM, which means that the same optical setting for SYBR Green I can also be used for EvaGreen^TM.

We now offer two EvaGreen^TM products: 1) Eva Green^TM at 20x concentration (#31000); and 2) EvaGreen^TM qPCR Basic Mix at 2x concentration (#31001). The Basic Mix contains everything you need to run a qPCR except for the Taq enzyme. We will soon offer EvaGreen^TM Qpcr Master Mix.

 

 

GelRed^TM is a superior red fluorescent nucleic acid dye specifically designed for both precast and post gel staining. It has a combination of desirable properties that no other commercial nucleic acid gel stains possess: high sensitivity, extraordinary stability, low toxicity and versatility.

Most of the current commercial gel stains are lacking in one or more aspects. For example, although ethidium bromide (EB), the most widely used nucleic acid gel stain, offers acceptable sensitivity in most of the cases, it is a highly mutagenic chemical and its use requires a destaining step to reduce background fluorescence. SYBR^TM Green I and SYBR^TM Gold have been promoted as the most sensitive gel stains by the manufacturer. However, SYBR^TM Green I and particularly SYBR^TM Gold degrade fairly rapidly under the slightly alkaline condition of the commonly used electrophoresis buffer or in the matrix of precast gels, resulting in unreliable gel staining.

GelRed^TM overcomes most of the drawbacks encountered by the current commercial gel stains. Not only does the dye offer superior detection sensitivity, but also exhibits remarkable stability and provides the flexibility of being used as either a precast gel stain (Figure 1) or a post gel stain (Figure 2). The dye has a major excitation peak at around 300 nm and a red emission at around 595 nm (See Figure 3). Thus the dye can be optimally excited with a common 300 nm UV transilluminator while its fluorescence emission is completely compatible with either a standard EB filter or a SYBR^TM filter (Figure 2). When used as a precast gel stain, GelRed can be microwaved with agarose or be subjected to other heating procedures commonly used in preparing EB precast gels. Unlike EB precast gels, however, GelRed precast gels have virtually no background fluorescence and are highly sensitive in detecting lowmolecular- weight DNA fragments. As with EB, precast gels made from GelRed^TM can be safely and conveniently stored for later use without compromising the performance of the gels, an utility SYBR dyes do not possess. When used as a post gel stain, GelRed completes the staining in as little as 30 minutes without the need for an extra destaining step. Moreover, since GelRed is hydrolytically stable under either acidic or alkaline condition, its 1X staining solution can be prepared in bulk for later use.

Equally important is the potentially improved safety of GelRed^TM over EB. Ames test was performed on both EB and GelRed^TM to compare frameshift mutagenicity using S.typhimurium TA98 by Litron Laboratories (200 Canal View Blvd. Ste.106, Rochester, NY 14623). Both dyes were tested in seven doses (0.6, 1.2, 3, 6, 12, 30 and 60 nmoles) in addition to control (0 nmoles) in the presence or absence of metabolic activation by rat liver S9 extract, each in duplicated plates. Dose-dependent increase of colony number of revertants indicates mutagenicity of the tested dyes and the colony number of revertants reflects the degree of mutagenicity. GelRed^TM causes no significant increase of colony number in the absence of metabolic activation in all seven doses tested, indicating lack of mutagenicity without metabolic activation. GelRed^TM causes dose-dependent increase of colony number in the presence of metabolic activation. However, the mutagenicity is greatly reduced in comparison to EB (detailed test results available upon request).

We offer GelRed^TM as a 10,000X concentrated solution in DMF (cat# 41000) for your flexibility and also for your convenience GelRed^TM 3X solution (cat#41001) that can be directly used for post gel staining. We will also soon offer ready-made GelRed^TM precast gels for the ultimate convenience.

FEATURES

• Superior Sensitivity

  The most sensitive and robust nucleic acid gel stain.

• Unsurpassed Thermal Stability, Hydrolytical Stability and Photostability

  Can be microwaved or subjected to other similar heating procedures for making agarose gels; stable in alkaline or acidic buffers at room temperature; highly photostable.

• Improved Safety

  Shown to be much less mutagenic than ethidium bromide by Ames test.

• Ultimate Flexibility

  Can be used for either precast or post gel staining; for either agarose gels or polyacrylamide gels; and for either dsDNA or ssDNA or RNA.

• Simple Staining Procedure

  Prepare and run precast gels as with EB without having to worry about dye stability; and takes as little as 30 minutes for post staining without the need for destaining.

• Minimal Effect on DNA Migration Pattern

  DNA migration pattern in GelRed precast gels similar to that in gels without dyes.

• No Need for Filter Change

  Works perfectly well with either a standard EB filter or a SYBR filter.

• Perfect Compatibility with a Standard 300 nm UV Transilluminator

  Maximally excited at around 300 nm UV(See Figure 3 for spectra)

The Most Sensitive and Stable Precast Gel Stain
		Figure 1. GelRedTM is significantly more sensitive than ethidium bromide (EB) for detecting low-level DNA, especially in the lower molecular weight area. As with EB, precast gels prepared from GelRedTM are stable for long-term storage, whereas precast gels made from SYBR Green I or GelStar degrade rapidly within a day. Shown above are dilutions of 1 Kb Plus DNA Ladder electrophoresed on 1% agarose gels precasted with GelRed or EB in 1x TBE. The total amount of DNA loaded per lane was: 200 ng, 100 ng, 50 ng and 25 ng from left to ight. Gels were imaged using 300-nm transillumination and photographed with an EB filter and Polaroid 667 black-and-white print films.

 

The Most Sensitive and Stable Post Gel Stain

Figure 2. GelRedTM displays consistently superior sensitivity for post gel staining, regardless of the filter used (A vs. C) and storage and handling condition. SYBR Gold, however, showed comparable performance only when used fresh from the manufacturer and with a SYBR filter (B vs. D). Following a few freeze-thaw cycles, SYBR Gold 10,000X solution degraded significantly, resulting in poor staining (E). SYBR Gold 1X solution also degrades (See Figure 4). Dilutions of 1 kb Plus DNA Ladder were electrophoresed on 1% agarose gels in 1x TBE and post- stained with GelRedTM (#41000 and #41001) and SYBR Gold, respectively. Gels were imaged using 300-nm transillumination and photographed with the indicated filters and Polaroid black-and-white print films. The total amount of DNA per lane for each serial dilution was: 200 ng, 100 ng, 50 ng and 25 ng from left to right.

Note: *GelRed and its uses are covered by pending US and international patents. **SYBR is trademark of Molecular Probes, Inc. and GelStar is trademark of FMC corporation.

Please also see our EvaGreen^TM(cat#31000), a breakthrough nucleic acid dye ideally suited for quantitative real-time PCR (qPCR). By incorporating a smart "release-on-demand" DNA-binding technology, EvaGreen^TM has low PCR inhibition while exhibiting superior sensitivity. Similar to our GelRed^TM, EvaGreen^TM has remarkable stability.

GelRed^TM is a superior red fluorescent nucleic acid dye specifically designed for both precast and post gel staining. It has a combination of desirable properties that no other commercial nucleic acid gel stains possess: high sensitivity, extraordinary stability, low toxicity and versatility.

Most of the current commercial gel stains are lacking in one or more aspects. For example, although ethidium bromide (EB), the most widely used nucleic acid gel stain, offers acceptable sensitivity in most of the cases, it is a highly mutagenic chemical and its use requires a destaining step to reduce background fluorescence. SYBR^TM Green I and SYBR^TM Gold have been promoted as the most sensitive gel stains by the manufacturer. However, SYBR^TM Green I and particularly SYBR^TM Gold degrade fairly rapidly under the slightly alkaline condition of the commonly used electrophoresis buffer or in the matrix of precast gels, resulting in unreliable gel staining.

GelRed^TM overcomes most of the drawbacks encountered by the current commercial gel stains. Not only does the dye offer superior detection sensitivity, but also exhibits remarkable stability and provides the flexibility of being used as either a precast gel stain (Figure 1) or a post gel stain (Figure 2). The dye has a major excitation peak at around 300 nm and a red emission at around 595 nm (See Figure 3). Thus the dye can be optimally excited with a common 300 nm UV transilluminator while its fluorescence emission is completely compatible with either a standard EB filter or a SYBR^TM filter (Figure 2). When used as a precast gel stain, GelRed can be microwaved with agarose or be subjected to other heating procedures commonly used in preparing EB precast gels. Unlike EB precast gels, however, GelRed^TM precast gels have virtually no background fluorescence and are highly sensitive in detecting lowmolecular- weight DNA fragments. As with EB, precast gels made from GelRed^TM can be safely and conveniently stored for later use without compromising the performance of the gels, an utility SYBR^TM dyes do not possess. When used as a post gel stain, GelRed^TM completes the staining in as little as 30 minutes without the need for an extra destaining step. Moreover, since GelRed^TM is hydrolytically stable under either acidic or alkaline condition, its 1X staining solution can be prepared in bulk for later use.

Equally important is the potentially improved safety of GelRed^TM over EB. Ames test was performed on both EB and GelRed^TM to compare frameshift mutagenicity using S.typhimurium TA98 by Litron Laboratories (200 Canal View Blvd. Ste.106, Rochester, NY 14623). Both dyes were tested in seven doses (0.6, 1.2, 3, 6, 12, 30 and 60 nmoles) in addition to control (0 nmoles) in the presence or absence of metabolic activation by rat liver S9 extract, each in duplicated plates. Dose-dependent increase of colony number of revertants indicates mutagenicity of the tested dyes and the colony number of revertants reflects the degree of mutagenicity. GelRed^TM causes no significant increase of colony number in the absence of metabolic activation in all seven doses tested, indicating lack of mutagenicity without metabolic activation. GelRed^TM causes dose-dependent increase of colony number in the presence of metabolic activation. However, the mutagenicity is greatly reduced in comparison to EB (detailed test results available upon request).

We offer GelRed^TM as a 10,000X concentrated solution in DMF (cat#41000) for your flexibility and also for your convenience GelRed^TM 3X solution (cat# 41001) that can be directly used for post gel staining. We will also soon offer ready-made GelRed^TM precast gels for the ultimate convenience.

FEATURES

• Superior Sensitivity

  The most sensitive and robust nucleic acid gel stain.

• Unsurpassed Thermal Stability, Hydrolytical Stability and Photostability

  Can be microwaved or subjected to other similar heating procedures for making agarose gels; stable in alkaline or acidic buffers at room temperature; highly photostable.

• Improved Safety

  Shown to be much less mutagenic than ethidium bromide by Ames test.

• Ultimate Flexibility

  Can be used for either precast or post gel staining; for either agarose gels or polyacrylamide gels; and for either dsDNA or ssDNA or RNA.

• Simple Staining Procedure

  Prepare and run precast gels as with EB without having to worry about dye stability; and takes as little as 30 minutes for post staining without the need for destaining.

• Minimal Effect on DNA Migration Pattern

  DNA migration pattern in GelRed precast gels similar to that in gels without dyes.

• No Need for Filter Change

  Works perfectly well with either a standard EB filter or a SYBR filter.

• Perfect Compatibility with a Standard 300 nm UV Transilluminator

  Maximally excited at around 300 nm UV(See Figure 3 for spectra)

The Most Sensitive and Stable Precast Gel Stain
		Figure 1. GelRedTM is significantly more sensitive than ethidium bromide (EB) for detecting low-level DNA, especially in the lower molecular weight area. As with EB, precast gels prepared from GelRedTM are stable for long-term storage, whereas precast gels made from SYBR Green I or GelStar degrade rapidly within a day. Shown above are dilutions of 1 Kb Plus DNA Ladder electrophoresed on 1% agarose gels precasted with GelRed or EB in 1x TBE. The total amount of DNA loaded per lane was: 200 ng, 100 ng, 50 ng and 25 ng from left to ight. Gels were imaged using 300-nm transillumination and photographed with an EB filter and Polaroid 667 black-and-white print films.

 

The Most Sensitive and Stable Post Gel Stain

Figure 2. GelRedTM displays consistently superior sensitivity for post gel staining, regardless of the filter used (A vs. C) and storage and handling condition. SYBR Gold, however, showed comparable performance only when used fresh from the manufacturer and with a SYBR? filter (B vs. D). Following a few freeze-thaw cycles, SYBR Gold 10,000X solution degraded significantly, resulting in poor staining (E). SYBR Gold 1X solution also degrades (See Figure 4). Dilutions of 1 kb Plus DNA Ladder were electrophoresed on 1% agarose gels in 1x TBE and post- stained with GelRedTM (#41000 and #41001) and SYBR Gold, respectively. Gels were imaged using 300-nm transillumination and photographed with the indicated filters and Polaroid black-and-white print films. The total amount of DNA per lane for each serial dilution was: 200 ng, 100 ng, 50 ng and 25 ng from left to right.

            

Note: *GelRed and its uses are covered by pending US and international patents. **SYBR is trademark of Molecular Probes, Inc. and GelStar is trademark of FMC corporation.