We offer a range of ultra pure, high-quality reagents for a variety of molecular biology applications such as nucleic acid electrophoresis, nucleic acid purification and sequencing, PCR and more.
Fisher BioReagents offers a wide range of reagents for Nucleic Acid Electrophoresis applications. Choosing the right reagents will allow you to achieve optimal results in your experiment. Use our quick selection guides to assist you in finding the right grades of Agarose, Buffer and Ladders.
Agarose is a linear polysaccharide composed of alternating residues of D- and L-galactose that are joined by glycosidic linkages. Agarose forms gels that are both porous and resilient. These gel properties provide a sieving matrix, which allows the electrophoretic separation of charged macromolecules, such as DNA or RNA, according to size. Compared to polyacrylamide gel, agarose has a lower resolution but wider range of separation. Lower grades of agarose can be contaminated with other polysaccharides, salts and proteins. Such impurities can alter the gelling/melting temperature of agarose solutions or affect the ability to use the recovered nucleic acid sample in a post-electrophoresis application.
Fisher BioReagents offers three different grades of agarose that are functionally tested and pre-qualified for specific applications. Agarose grades used in electrophoresis of nucleic acids:
1. Size of DNA or RNA fragments to be analyzed:
|Agarose||Agarose Separation Ranges|
|Low EEO/Multipurpose||500bp to 23kb|
|Low Melting/Nucleic Acid Recovery||200bp to 25kb|
|Broad Separation Range for DNA/RNA||500bp to 25kb|
|Low Melting <1kb DNA/RNA||50bp to 1kb|
|Intermediate Melting||15bp to 1.2kb|
2. Type of downstream application that will follow electrophoretic separation (eg. cloning procedures directly from re-melted agarose or in-gel reaction).
|Type of Agarose||Low EEO||Low Melting > 200bp||Low Melting > 1000bp||Wide Separation Range||PCR Grade|
|Recovery of DNA and RNA||x||x||x||x||x|
|Southern and Northern Biots||x|
|DNA/RNA Separation 50bp to 1kb||x||x|
|DNA/RNA Separation > 1kb||x||x||x|
|PCR Fragment Analysis||x||x||x||x||x|
|In-gel Reactions (ligation, transformations, PCR)||x|
|Available Pack Sizes||100g to 500g||25g||100g||100g to 500g||100g|
|Agarose Grade||Molecular Biology||Molecular Biology||Genetic Analysis||Genetic Analysis||PCR|
Two buffers commonly used for DNA agarose electrophoresis are Tris-acetate with EDTA (TAE; 40mM Tris-acetate, 1mM EDTA) and Tris-borate with EDTA (TBE, 89mM Tris-borate, 2mM EDTA). Because the pH of these buffers is neutral, the phosphate backbone of DNA has a net negative charge and migrates toward the anode. TAE and TBE have different properties, which makes one more suitable than the other for a specific purpose.
The denaturing system chosen depends on the purpose of the RNA experiment and the size of the RNA fragment being separated. Formaldehyde denaturation is suitable if RNA samples are to be recovered. Formamide denaturation is suitable if the RNA needs to retain its biological activity.
|TAE||DNA recovery; Electrophoresis of large DNA (>12 kb)||Low buffering capacity; recirculation may be necessary for extended run times (>6 hr)|
|TBE||Electrophoresis of small DNA (<1 kb); Increased resolution of small DNA (< 1kb)||Decreased DNA mobility; High buffering capacity - no recirculation required for extended run times|
|MOPS||Electrophoresis of formaldehyde denatured RNA||Buffer is low in ionic strength; recirculation of buffer may be necessary|
Suggested Agarose Concentrations
The optimal gel concentration depends on the size of the DNA fragments to be resolved.
|Product Description||Main Application||DNA Size Range in Base Pairs||Final Agarose Concentation % (W/V) 1x TAE Buffer||Final Agarose Concentration % (W/V) 1x TBE Buffer|
Low Melting, <1kb
|Certified recovery of small
nucleic acid fragments,
Low Melting, Nucleic<1kb
|Broad separation range,
Ideal for DNA and RNA
Range for DNA/RNA,
|Suitable for routine nucleic
applications with broad
|Routine electrophoresis of
DNA and RNA. High gel
strength ideal for Southern
and Northern blotting.
To achieve the most accurate qualitative and quantitative analysis via agarose gel electrophoresis, the appropriate DNA or RNA standard is required. Fisher BioReagents provides a wide range of standards, including routine DNA ladders for quick size and quality assessment as well as Fisher Bioreagents™ exACTGene™ DNA ladders that allow for quantitative analysis.
Fisher Bioreagents™ RiboLadders™ RNA Standards
These standards can be used to assess single-stranded RNA molecules on both native and denaturing agarose gels. These unique RNA standards are lyophilized to reduce thawing-related degradation, to prolong shelf life and to ensure consistent performance.
|Application||Size Range||Number of Bands||Number of Loadings|
|Small RNA fragments||0.1-1kb||8||50|
|Large RNA fragments||0.2-4kb||9||50|
Routine DNA Ladders/RiboLadders™ RNA Standards
exACTGene and Routine DNA Ladders
Ready-to-use (pre-mixed with the loading dye), room temperature, stable DNA ladders are available for all common electrophoresis applications.
|Application||Size Range||Numbers of Bands||Number of Loadings|
|exACTGene DNA ladders are ideal for qualitative analysis, quantitative estimation, and size assessment|
|PCR fragment analysis||25-650bp||14||100-10uL|
|PCR fragment analysis, small DNA digests||25-1,000bp||12||100/10uL|
|Quick check of PCR or enzyme digestion results||50-2,000bp||8||100/10uL|
|General purpose, small DNA fragments||100-1,000bp||10||100/10uL|
|Fast run times, small DNA fragments||100-2,000bp||8||100/10uL|
|Large size PCR or cloning||100-2,000bp||10||100/10uL|
|Small or large cloning application||100-5,000bp||16||100/10uL|
|General purpose, large digested DNA||300-10,000bp||13||100/10uL|
|General purpose, wide size range||100/10,000bp||19||100/10uL|
|General purpose, extra-large fragments||300-24,000bp||15||100/10uL|
|Routine DNA ladders are designed for qualitative analysis and size assessment|
|small fragments, quick size assessment||50-2,000bp||11||200/5uL|
|Quick size assessment of broad size range||50-10,000bp||16||200/5uL|
exACTGene DNA Ladders: Routine DNA Ladders/RiboLadders™ RNA Standards