Lonza Rockland, Inc.
www.lonza.com
scientific.support@lonza.com
Scientific Support: 800-521-0390
Customer Service: 800-638-8174
Document # 18111-0612-09
Rockland, ME 04841 USA

GelStar™ Nucleic Acid Gel Stain
Instructions for Use
Introduction
GelStar™ Stain is a highly sensitive fluorescent stain for
detecting nucleic acids in agarose and polyacrylamidegels. This single stain gives high sensitivity detection of
double-stranded or single-stranded DNA and RNA.
Gels can be post-stained or alternatively the stain can beadded to agarose gels during gel casting. A comparisonof GelStar™ Stain with ethidium bromide is shown
below. These results were obtained using a 300 nm UV
light source and the appropriate filter for each stain.
Nucleic AciddsDNA
ssDNA or SSCP DNA
Glyoxalated RNA
Native RNA
Formaldehydedenatured RNA

upon the number and size of gels stained and theamount of nucleic acid in the gels.
Cautions• The DMSO stock solution should be handled withcaution as DMSO is known to facilitate entry oforganic molecules into tissues. We strongly
recommend using double gloves when handling theDMSO stock solutions. Please refer to theGelStar™ Stain MSDS for more details. No data areavailable on the mutagenicity or toxicity of theGelStar™ Stain.
• This stain should be treated as a potential
mutagen and used with appropriate care. Stain
solutions should be disposed of by passing
through activated charcoal followed byincineration of the charcoal. Solutions may also
be treated using commercially available units
designed to bind ethidium bromide.

Sensitivity Increase
Compared toEthidium Bromide
4-16 fold20-80 fold
16 fold
3-10 fold
2-3 fold

Storage and Handling• GelStar™ Stain is supplied as a 10,000X
concentrated stock solution in DMSO. Store
desiccated at -20°C protected from light. When
stored per directions the stock solution is stable for
12 months from the date of receipt.
• Allow time for the stock solution to thaw totally.
Removal of stain from partially thawed solutions will
result in depletion of stain over time.
• Prepare and store the stain in polypropylene
containers such as Rubbermaid® Containers or
pipette-tip box lids. The stain may adsorb to glass
surfaces and some plastic surfaces, particularly if
the surfaces carry residues of anionic detergents orreagents.
• GelStar™ Stain may be diluted in most common
electrophoresis buffers with a pH range from 7.0-8.5
or in TE buffer. Staining solutions prepared in wateror in buffer with a pH below 7.0 or above 8.5 areless stable and show reduced staining efficiency.
• Diluted stock solutions are stable for several days
when stored at 4°C in a polypropylene container
protected from light. The number of reuses depends

Application Notes• The powder used on some laboratory gloves may
contribute to background fluorescence. We
recommend the use of powder-free gloves andrinsing gloves prior to handling gels.
• Fibers shed from clothing or lab coats may be
fluorescent; be cautious when handling gels.
• Staining of nucleic acids with GelStar™ Stain has
minimal impact on blotting efficiency. To ensure
efficient hybridization, use of prehybridization andhybridization solutions containing 0.1%-0.3% SDS isimportant to remove stain retained during transfer.
• GelStar™ Stain can be removed from nucleic acids
by ethanol precipitation. Isopropyl alcohol
precipitation is less effective at removing the dye;
butyl alcohol extraction, chloroform extraction andphenol do not remove the dye efficiently.

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Before You Begin
Detection of DNA and RNA in Agarose gels byPost-Staining
1. Separate the samples by electrophoresis as normal.
2. Remove GelStar™ Stain from -20°C storage andthaw at room temperature for 10 to 20 minutes.
3. Spin the stain vial briefly in a microcentrifuge todeposit the solution in the bottom of the vial.
4. Dilute stock GelStar™ Stain in buffer in a
polypropylene container.

Use a polypropylene container for diluting the stainand staining the gel
Use a 302 or 312 nm UV transilluminator or a bluelight transilluminator such as the Clare ChemicalDark Reader® Transilluminator for dye excitation
Use a GelStar™ Filter for photography

For DNA:
Dilute in TE, TAE, or TBE
to give a final
concentration of 1X, e.g.,
5 µl of stain stock added
to each 50 ml of buffer

Detection of DNA or RNA in Agarose Gels byPre-staining
1. Remove GelStar™ Stain from -20°C storage andthaw at room temperature for 10 to 20 minutes.
2. Spin the stain vial briefly in a microcentrifuge todeposit the solution in the bottom of the vial.
3. Prepare the gel solution and cool the agarose
solution to 55°C-65°C.
4. Add a volume of stock GelStar™ Stain to thetempered gel solution as indicated below:
For DNA:For RNA:

Use a final concentrationof 1X e.g., 5 µl of stain
stock added to each
50 ml of gel solution.

Use a final concentrationof 2X, e.g., 10 µl of stain
stock added to each 50 mlof gel solution.

For RNA:
Dilute stock GelStar™ Stain in 1X
MOPS buffer to give a final
concentration of 2X, e.g., 10 µl ofstain stock added to each 50 ml ofbuffer.

5. Prepare enough stain solution to cover the gel
during staining.
6. Mix the stain solution to distribute the stain
thoroughly into the solution.
7. Place the gel and the stain solution in a
polypropylene container and incubate with gentle
agitation. Protect from direct exposure to strong lightduring staining. Staining is normally complete within
30 minutes. Exceptionally thick gels (>4 mm) or highconcentration gels may require longer staining times
for optimal results.
8. Visualize the results by photography, scanning, orimage capture as described, on next page. No
destaining is normally required, although werecommend a brief rinse with buffer to minimize
deposition of stain on work surfaces.

5. Mix the gel solution by swirling, stirring, or inversion
to thoroughly distribute the stain into the gel solution.
6. Immediately cast the gel and allow to solidify.
Avoid extended light exposure and extendedexposure (>10 min.) to a buffer overlay.
7. Run the gel using your standard protocol.
8. Visualize the results by photography, scanning, orimage capture as described, on next page. No
destaining is normally required, although we
recommend a brief rinse with buffer to minimize
deposition of stain on work surfaces.

NOTES:• GelStar™ Stain is compatible with post-staining ofDNA in agarose gels >4 mm thick, such as Lonza
Reliant™ Precast Agarose Gels.
• GelStar™ Stain gives excellent detection of RNA
that has not been denatured, as well as RNA
denatured by a variety of methods, e.g., glyoxal,formamide, or formaldehyde. The increase indetection sensitivity in comparison to ethidium
bromide staining varies depending upon the sampleand gel preparation methods used (see note belowand Table on page 1).
• Detection of glyoxalated RNA with GelStar™ Stain isoptimal in gels that are ≤4 mm thick. To see the fullsensitivity enhancement of GelStar™ Stain, use gelsof this thickness or include the stain in the gel fordetection.

NOTES:• The effect of GelStar™ Stain on DNA migration issmaller compared to ethidium bromide, i.e., DNAmigration is slower compared to gels with no stain
added.
• GelStar™ Stain is sensitive to the presence ofparticulates in the gel buffer and dust/debris on geltrays. For optimal results, filter the buffer used toprepare the gel solution.
• Including GelStar™ Stain in vertical gels is notrecommended as the dye may bind to glass orplastic plates.

2Excitation of GelStar™ Stained Nucleic Acids

Detection of Nucleic Acids on Vertical Gels by PostStaining
1. Separate the samples by electrophoresis as normal.
2. Remove GelStar™ Stain from -20°C storage andthaw at room temperature for 10 to 20 minutes.
3. Spin the stain vial briefly in a microcentrifuge todeposit the solution in the bottom of the vial.
4. Add stock GelStar™ Stain to buffer (TE, TAE,MOPS, or TBE) to give a final concentration of 1X,e.g., 5 µl of stain stock added to each 50 ml ofbuffer. Prepare enough stain solution to cover thesurface of the gel during staining.
5. Mix the stain solution by swirling, stirring, orinversion to distribute the stain thoroughly into thesolution.
6. Open the cassette, and leave the gel in place on oneplate.
7. Place the plate, gel side up, in a staining container.
8. Gently pour the stain over the surface of the gel; adisposable pipette may be used to help distribute thestain evenly over the gel surface. Do not submerge
the gel and plate in staining solution.
9. Incubate for 30 minutes. No destaining is required,although we recommend a brief rinse with buffer tominimize deposition of stain on work surfaces.
10. Visualize the results by photography, scanning, orimage capture as described above. For highest
sensitivity the gel should be carefully removed fromthe plate and placed directly on the transilluminatoror scanning stage. Alternatively, if a relatively low
fluorescence plate is used, the results may be
visualized by placing the gel and plate gel side downon the transilluminator and photographing or byscanning the gel directly on the plate.

Either:Illuminate the gel with a standard UV transilluminator(302 or 312 nm).
Or:
Illuminate the gel with a blue-light transilluminator suchas the Clare Chemical Dark Reader® Transilluminator .
Or:
Excite the GelStar™ Stain with an argon laser scanningsystem. Systems compatible with the detection ofSYBR® Green Stain should also be compatible withGelStar™ Stain.
Visualization by Photography
Photograph the gel with the filter and film in the tablebelow. Polaroid® type 55 positive/negative film can beused for photography of gels having relatively strong
signals.
Exposure time varies with the strength of theillumination source and the filter used for photography.
Suggested Exposure Conditions For Different Film
Types.
Film
Type 57
or 667
Type 55

f-stop

FilterExposure

4.5

GelStar™ Filter

2-5 seconds

4.5

GelStar™ Filter

15-45
seconds

Visualization by Image Capture systems
GelStar™ Stain is compatible with most CCD and video
imaging systems. Due to variations in the filters forthese systems you may need to purchase a new filter.
Lonza does not sell filters for this type of camera.
Contact your systems manufacturer and using theexcitation and emission information listed they can guide
you to an appropriate filter. The excitation and emission
maxima of GelStar™ Stain are 493 nm and 527 nm (532nm for RNA) respectively.

NOTES:• As an alternative to the protocol presented forstaining gels on the cassette plate, smaller gels suchas minigels may be removed from both plates thenstained using the protocol for post-staining agarosegels.
• Treatment of one plate with a “release” agent, suchas Gel Slick™ Solution (Lonza Catalog No. 50640),
increases the ease of separation the glass plates
while keeping the gel in place on the other plate forstaining.
• Handling or compression of gels (particularly
polyacrylamide-type gels) can lead to regions of highbackground after staining. If possible gels should
not be handled directly; use a spatula (or other tool)
and a squirt bottle to slide the gel off the plates andinto the stain or onto the light box.
• Addition of 50% glycerol to the staining buffer isrecommended when using GelStar™ Stain with
MDE™ Gels (Lonza Catalog No. 50620) for
heteroduplex or SSCP analyses. This minimizes
swelling of the gel during staining, and improves gel
handling and staining intensity.

3Ordering Information
Catalog
No.
50535
50536

Description

Quantity

GelStar™ Nucleic Acid Stain
GelStar™ Nucleic Acid Stain
Photographic Filter2 x 250 µl
1 eachRelated products
DNA Ladders
MetaPhor™ Agarose
NuSieve™ GTG™ Agarose
NuSieve™ 3:1 Agarose
PAGEr™ Gold TBE Precast Gels
SeaKem® GTG™ AgaroseSeaKem® LE Agarose
SeaPlaque™ GTG™ Agarose
MDE™ Gel Solution
For more information contact Scientific Support at(800) 521-0390 or visit our website at www.lonza.com
GelStar™ Stain is manufactured for Lonza Rockland,Inc.
For Research Use Only.
Not for use in diagnostic procedures.

SeaKem and GelStar are trademarks of FMC Corporation,
Rubbermaid is a trademark of Rubbermaid, inc., Polaroid is atrademark of Polaroid Corp., SYBR is a trademark of MolecularProbes, Inc. All other trademarks herein are marks of the Lonza Groupor its affiliates. GelStar Stain is covered by U.S. Patent 5,436,134.
Other U.S. and foreign patents pending.
 2012 Lonza Rockland, Inc.
All rights reserved.

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