Nucleic Acid Electrophoresis Everything for convenient, accurate nucleic acid analysis— except the sample Nucleic Acid Electrophoresis The most complete collection of nucleic acid electrophoresis and analysis tools → Easy ordering from an all-inclusive supplier → Ready-to-use products reduce opportunities for introducing error → Thoroughly tested reagents provide proven results Simplify your life. Instead of running around placing and tracking multiple orders, call Invitrogen, your one-stop shop. You’ll find all the high-quality tools you need for dependable, accurate performance in your nucleic acid electrophoresis and analysis experiments. In this guide you’ll find a brief overview of the tools available. For more information, visit the online resources listed in Table 1. Tools Page Precast, hassle-free agarose gels 3 Safe, sensitive DNA stains 10 Convenient nucleic acid markers 11 Highest purity and reliability with UltraPure™ reagents 13 Table 1—Online resources. Description Website E-Gel® precast agarose gels www.invitrogen.com/egels DNA markers www.invitrogen.com/namarkers SYBR® Safe DNA Gel Stain www.invitrogen.com/sybrsafe Safe Imager™ Blue-Light Transilluminator www.invitrogen.com/safeimager UltraPure™ reagents www.invitrogen.com (search for “UltraPure”) Protein electrophoresis www.invitrogen.com/proteomics Ordering www.invitrogen.com Precast, hassle-free agarose gels E-Gels make agarose electrophoresis fast, easy, and safe Each E-Gel® agarose gel is a self-contained, complete electrophoresis system that includes agarose, your choice of DNA gel stain (or no gel stain), electrodes, and patented ion-exchange matrices contained inside a disposable, UV-transparent cassette. With E-Gels, you don’t have to pour gels, make buffer, or even have a gel box and power supply. Just load your samples and run (Figure 1). You’ll have results in as little as 7 minutes. E-Gels are available in a variety of agarose percentages and formats to meet all your throughput needs. And they are stable at room temperature so you can keep them on your bench, always ready to run. E-Gels run in their own bases that include a built-in power supply. Choose either the new E-Gel® iBase™ Power System (page 6) or the original E-Gel® PowerBase™ v.4 (page 7). 1. Remove E-Gel® cassette from pouch. Innovative, patented technology E-Gel® cassettes contain two unique ion-exchange matrices (IEMs) that are in contact with the running gel and electrodes (Figure 2). The IEMs supply a continuous flow of ions throughout the gel. This patented technology results in a sustained electrical field with enhanced buffering capacity that eliminates the need for liquid buffer and exposure to ethidium bromide. 2. Insert into E-Gel® iBase™ (shown here) or E-Gel® PowerBase™ unit. U. S. P. 5 58 2 8 7 6 5 3 2 Upper IEM The upper IEM, near the cathode, contains acetate anions. AC– Upper IEM OH– Ac– EG el ™ 1 A ga 4 ro se (G P) Cathode (–) 70 2 9 10 11 12 Running gel Tris+ Anode (+) Lower IEM H+ Lower IEM The lower IEM, near the anode, contains Tris cations and ethidium bromide. Tris+ Figure 2—Location of the IEMs in an E-Gel® cassette. Protons and hydroxyl ions, by-products of water electrolyzed during electrophoresis, are exchanged with Tris, ethidium, and acetate ions in the IEMs. 3. Load samples and run. Get results in 7–30 minutes. Figure 1—Using E-Gels is as easy as 1-2-3. www.invitrogen.com Nucleic Acid Electrophoresis Single- and double-comb E-Gels for lowthroughput applications A B Get superior resolution in low-throughput agarose electrophoresis with E-Gels. Single-comb E-Gels contain a single row of 12 sample wells. Double-comb E-Gels contain two rows, each with eight sample wells and one marker well. The wells are spaced Figure 3—Sample loading using single- (A) and double-comb (B) E-Gels (in the E-Gel® PowerBase™ v.4). for compatibility with 8-channel pipettors for easy, multi-sample loading (Figure 3). Single- and double-comb E-Gels can be run in either the new E-Gel® iBase™ Power System or the original E-Gel® A B2% Agarose 2% Agarose (GP) (GP) 0.8% 0.8% Agarose Agarose (GP) (GP) 1 2 13 24 35 46 57 6 8 97 10 8 911 10 12 11 12 PowerBase™ v.4 (pages 7–8). 4% Agarose 4% Agarose (HR) (HR) 1 2 13 24 35 46 57 68 79 810 911 10 12 11 12 1 2 13 24 35 46 57 68 E-Gels are offered in a variety of agarose concentrations (Figures 4 and 5). Choose the gel percentage that best suits your needs (Table 2). E-Gels come with a convenient selection of DNA staining options. Choose E-Gels with new 0.8% SYBR® Safe DNA Gel Stain or Agarose (GP) 2% Agarose (GP) 1 2 3 4 5 6 7 8 9 10 11 12 1 2 ethidium bromide incorporated, or Clear E-Gels containing no 3 4 5 6 C 7 8 9 10 11 12 4% Agarose (HR) 1 2 3 4 5 6 7 8 9 10 11 12 DNA stain (Figure 6). 100 bp 50 bp A B 0.8% Agarose (GP)–Double Comb 1 2 3 4 M 5 6 7 2% Agarose (GP)–Double Comb 8 1 2 3 4 M 5 6 7 8 10 kb 500 bp 1 kb 100 bp 10 kb 500 bp 1 kb 9 10 11 12 M 13 14 15 16 100 bp 9 10 11 12 M 13 14 15 16 Figure 5—Electrophoresis results using double-comb E-Gels. A. 0.8% agarose. Lanes 1, 8, 9, 16: High DNA Mass Ladder. Lanes 2, 10: pcDNA™3.1 Nco I-cut. Lanes 3, 11: pcDNA™3.1 uncut. Lanes 4, 12: pUC18 Pst I-cut. M: Low DNA Mass Ladder. Lanes 5, 13: 1 kb PCR product. Lanes 6, 14: 3 kb PCR product. Lanes 7, 15: 5 kb PCR product. B. 2% agarose. Lanes 1, 8, 9, 16: 1 Kb Plus DNA Ladder. Lanes 2, 10: pcDNA™3.1 Nco I-cut. Lanes 3, 11: pcDNA™3.1 uncut. Lanes 4, 12: pUC18 Pst I-cut. M: Low DNA Mass Ladder. Lanes 5, 13: 500 bp PCR product. Lanes 6, 14: 1 kb PCR product. Lanes 7, 15: 2 kb PCR product. Figure 4—Electrophoresis results using single-comb E-Gels. A. 0.8% agarose with ethidium bromide. Lane 4: 1 Kb Plus DNA Ladder. Lanes 6–9: 3,871 bp PCR product amplified with Pfx50™ DNA Polymerase. B. 2% agarose with ethidium bromide. Lane 4: 1 Kb Plus DNA Ladder. Lanes 6–9: 3,123 bp PCR product amplified with Pfx50™ DNA Polymerase. C. 4% agarose with ethidium bromide. Lanes 1, 2, 11, 12: 50 bp DNA Ladder. Lanes 5, 6: 25 bp DNA Ladder. Lanes 7, 8: 10 bp DNA Ladder. Lanes 3, 9: 450 bp PCR product. Lanes 4, 10: 500 bp PCR product. Table 2—E-Gels come in a range of agarose percentages. Gel percentage Resolution range 0.8% 800 bp to 10 kb 1.2% 100 bp to 5 kb 2% 100 bp to 2 kb 4% 20 bp to 500 bp 1.2% Agarose (GP) 1 2 3 4 5 6 7 8 9 10 11 12 E-Gel with SYBR® Safe ® A B 1.2% Agarose 1.2% Agarose (GP) (GP) 1.2% Agarose 1.2% Agarose (GP) (GP) 8 11 9 12 10 11 12 1 2 1 2 31 24 3 5 46 57 86 97 10 13 24 35 46 57 68 79 810 9 11 10 12 11 12 1 2 1 3 2 4 3 5 4 6 5 7 6 8 7 9 8 10 9 1110 12 11 A 12 B 2% Agarose (GP) 1.2% Agarose (GP) 1 2 3 E-Gel® with ethidium bromide 4 5 6 4 5 6 7 8 9 10 11 12 E-Gel® with SYBR® Safe DNA Gel Stain 7 8 9 10 11 12 1 2 3 4 5 6 7 8 9 10 11 12 E-Gel with SYBR® Safe ® Clear E-Gel® Figure 6—A selection of E-Gel® precast gels to suit your staining preferences. A. 1.2% agarose with ethidium bromide, viewed on a standard UV transilluminator. B. 1.2% agarose with SYBR® Safe stain, viewed on the Safe Imager™ Blue-Light Transilluminator. C. 1.2% Clear E-Gel® stained with 1:10,000 dilution of SYBR® Gold stain for 40 minutes and viewed on the Safe Imager™ Blue-Light Transilluminator. All gels were loaded with identical samples. Lane 3: 1 Kb Plus DNA Ladder. Lane 5: 462 bp PCR product. Lane 6: 2,497 bp PCR product. Lane 7: 3,123 bp PCR product. All PCR products were amplified with Pfx50™ DNA Polymerase. Lane 8: 3,871 bp PCR product. Lane 11: Low DNA Mass Ladder. E-Gel with SYBR® Safe ® Figure 7—Clear, easy-to-see bands with SYBR® Safe DNA Gel Stain. Gels were visualized on a standard 2% Agarose (GP) 300 nm UV transilluminator. Photographs were taken using1 the MiniBis photo documentation system from DNR, and the SYBR® 2 3 4 5 6 7 8 9 10 11 12 Safe photographic filter using an exposure time of 1.8 sec. A. E-Gel® 1.2% with SYBR® Safe stain. Lanes 1, 12: High DNA Mass Ladder. Lanes 2, 11: High Range DNA Ladder. Lane 3: 1 kb PCR product. Lane 4: 3 kb PCR product. Lane 5: 9 kb PCR product. Lanes 6, 7: 1 Kb Plus DNA Ladder. Lane 8: pBR322 EcoR I-cut. Lane 9: pBR322 uncut. Lane 10: pUC19 EcoR I-cut. B. E-Gel® 2% with SYBR® Safe stain. Lanes 1, 12: Low DNA Mass Ladder. Lanes 2, 11: E-Gel® Low Range Quantitative DNA Ladder. Lane 3: 240 bp PCR product. Lane 4: 317 bp PCR product. Lane 5: 1 kb PCR product. Lanes 6, 8: 100 bp DNA Ladder. Lane 7: 50 bp DNA Ladder. Lane 9: pUC19 EcoR I-cut. Lane 10: pUC19 uncut. E-Gel with SYBR® Safe ® 10,000 E-Gel® precast gels with SYBR® Safe DNA stain for enhanced safety SYBR® Safe stain/blue light 1,000 EtBr/UV CFU 3 1 2 3 6 7 8 9 10 11 12 C 1.2% Agarose (GP) 1 2 4 5 100 Enjoy the benefits of E-Gel® technology and avoid working with ethidium bromide. E-Gel® pre-cast gels with SYBR® Safe DNA stain are run using the same easy protocol as the original E-Gels. Directly visualize your DNA using an imaging system such as a laser-based scanner, , or a standard UV transilluminator (Figure 7). For even greater safety, use the Safe Imager™ Blue-Light Transilluminator (Figure 8). By eliminating ethidium bromide and UV transillumination, you enjoy better personal safety and obtain better cloning efficiency by preventing DNA damage. 10 0 15 30 45 60 120 Exposure time (seconds) Figure 8—Even a 30 second exposure to ethidium bromide and UV reduces cloning efficiency. Gels loaded with equal amounts of a PCR product (1.25 kb gene fragment from Ultimate™ ORF IOH# 11050) were stained with either SYBR® Safe DNA Gel Stain (1:10,000 in TBE) or ethidium bromide (0.5 µg/ml in TBE) following electrophoresis. The gels were visualized on either a UV transilluminator (ethidium bromide) or blue-light box (SYBR® Safe stain), and bands excised after defined exposure times. DNA was purified from the gel fragments under identical conditions and used in parallel subcloning reactions. Following transformation into One Shot® TOP10 Chemically Competent E. coli, three serial dilutions were plated and colonies counted using an Alpha Innotech imaging system. www.invitrogen.com Nucleic Acid Electrophoresis The easiest way to extract your DNA band: E-Gel® CloneWell 0.8% SYBR® Safe gels and the E-Gel® iBase™ Power System If cloning is your application, you need E-Gel® CloneWell 0.8% SYBR® Safe gels and the E-Gel® iBase™ Power System. Using them together, you’ll be able to extract your DNA bands of interest from the gel without any additional purification steps, saving you time and effort while maximizing the amount of DNA you recover. Gels that are smarter by design The CloneWell gel features two rows of wells (Figure 9). To extract your DNA band of interest without gel purification, simply load your sample into the top row and electrophorese until your band moves into the second row of wells. Then use a pipette to easily remove your purified DNA band. Guides on the gel cassette help you track your band of interest so you know when your DNA is ready for retrieval. There is no need to cut a gel slice and risk losing your DNA during a gel purification step. The 0.8% agarose concentration is ideal for separating bands up to 5 kb in length. SYBR® Safe DNA Gel Stain is incorporated into the gel itself, allowing you to visually monitor migration on a Safe Imager™ Blue-Light Transilluminator. A base designed for tracking Figure 9—The E-Gel® CloneWell 0.8% SYBR® Safe gel. The two rows of wells allow you to load samples in the top row and capture the separated bands in the bottom row. The new E-Gel® iBase™ Power System, like the original E-Gel® PowerBase™ unit, is a selfcontained device that includes a built-in power supply. Simply connect the AC adaptor provided and plug into an electrical outlet. The E-Gel® iBase™ system includes upgraded features that give you faster run times and better control: the easy-to-read LCD display lets you view program selection and running time; preset programs are available for various gel types, or you can manually set your own run times; a reverse program allows you to change the field direction, ensuring complete capture of DNA when using new CloneWell gels; and a speed setting allows you to separate DNA in just 7 minutes.. In addition, the E-Gel® iBase™ Power System includes an automatic shutoff feature, so you won’t overrun your gel. The E-Gel® iBase™ Power System fits on a Safe Imager™ Blue-Light Transilluminator (Figure 10) so you can safely observe the DNA bands migrate in the gel. Figure 10—For maximum convenience and safety, run E-Gel® CloneWell gels in the E-Gel® iBase™ Power System on a Safe Imager™ Blue-Light Transilluminator. The most economical way to get started E-Gels are available in trial-size Starter Paks, each containing six single-comb E-Gels in 0.8%, 1.2%, 2%, or 4% agarose and an original E-Gel® PowerBase™ v.4 (Figure 11). All you need are your samples and you’re ready to experience E-Gel® convenience. The E-Gel® PowerBase™ v.4 simplifies electrophoresis because it’s a base and power supply all in one (Figure 12). To use it, simply place your E-Gel® cassette into the PowerBase™ unit, attach the AC adaptor provided, plug into the nearest electrical outlet, and run. The E-Gel® PowerBase™ v.4 has a built-in timer, alarm, and three-color display that indicates when specific programs begin and end. The automatic shutoff prevents gels from ever overrunning. Medium-throughput E-Gel® 48 gels for automation flexibility E-Gel® 48 gels are ready-to-use, 48-well agarose gels designed for medium-throughput analysis of PCR products and DNA fragments. Each gel contains 48 sample wells and four marker wells with a 3.2 cm run length. Load E-Gel® 48 gels with multichannel pipettors or with automated liquid handling systems* for increased throughput (Figure 13). DNA bands Figure 13—Loading an E-Gel® 48 gel using a liquid handling robot. resolve clearly in just 20 minutes (Figure 14). * Robotic scripts for loading E-Gel® 48 gels with various liquid handling systems are available on the Invitrogen website at www.invitrogen.com/egels. Figure 11—The convenient E-Gel® Starter Pak. The E-Gel® Starter Pak contains everything you need to start running E-Gels: six single-comb E-Gels in one of four agarose concentrations, and an E-Gel® PowerBase™ v.4. Figure 12—The E-Gel® PowerBase™ v.4. Just attach the AC adaptor provided to the E-Gel® PowerBase™ v.4, plug into an electrical outlet, and you are ready to run E-Gels. Figure 14—Clear electrophoretic resolution on a 4% E-Gel® 48 gel. Lanes 1, 20–21, 38–39, 48, M (lower right): 25 bp DNA Ladder. Lanes 2–3: Synthetic 21-mer siRNA. Lanes 4–5: undiced dsRNA. Lanes 6–7: diced dsRNA. Lanes 8, 13–14, 34–35: E-Gel® Low Range Quantitative DNA Ladder. Lanes 9–12, 15–19, 28–33, 36–37, 40–45: various PCR products (restriction enzyme digested). Lane 22: dsDNA, 60 bp. Lane 23: ssDNA. Lanes 24–25, M (lower left): 50 bp DNA Ladder. Lanes 26–27, M (upper right): 100 bp DNA Ladder. Lanes 46–47, M (upper left): 10 bp DNA Ladder. www.invitrogen.com Nucleic Acid Electrophoresis E-Gel® 96—a breakthrough in high-throughput electrophoresis E-Gel® 96 gels are designed for fast, high-throughput DNA electrophoresis. Each gel contains 96 sample wells and eight marker wells with a 1.6 cm run length, in a patented staggered-well format that is compatible with 8-, 12-, and 96-tip automated loading robots (Figure 15), to provide optimum performance and resolution in high-throughput applications† (Figure 16). With just 12-minute run times, up to 20,000 samples can be run in a single day, saving you weeks of screening time. For precise and effortless tracking of gels, each cassette is individually labeled with an EAN39 bar code that is recognized by most commercially available robotic bar code readers. † Robotic scripts for loading E-Gel® 96 gels with various liquid handling systems are available on the Invitrogen website at www.invitrogen.com/egels. Integrated power supplies for fast, automated electrophoresis E-Gel® 48 and E-Gel® 96 gels run on a specially designed, space-saving system of Mother and Daughter E-Bases—each a combined gel base and power supply all in one (Figure 17). These integrated, all-inclusive power supplies have a small benchtop footprint measuring only 5.5 × 6 inches per base, and can be plugged directly into any standard electrical outlet. Con- Figure 15—Loading an E-Gel® 96 gel using an automated liquid handling robot. nect multiple Daughter E-Bases to the Mother E-Base™ unit to create a multi-unit system capable of running over twenty gels simultaneously in just 12 minutes. Fluorescent EAN39 bar code Figure 17—The Mother E-Base™ and Daughter E-Base™ combination. Figure 16—Electrophoretic resolution on a 2% E-Gel® 96 gel. Lanes 1–3: 1 kb PCR product. Lanes 4–6: pcDNA™ 3.1 Nco I-cut. Lanes 7–9: 2 kb PCR product. Lanes 10–12: 500 bp PCR product. Lane M: E-Gel® Low Range Quantitative DNA Ladder. Load E-Gel® 48 and 96 gels robotically on the E-Holder™ Platform The handy E-Holder™ Platform is used to keep E-Gel® 48 and 96 gels stable during robotic loading of multiple gels (Figure 18). Designed for compatibility with the SBS (Society for Biomolecular Screening) standard-plate format, the E-Holder™ Platform’s spring-loaded mechanism ensures that each gel stays securely in place when the robotic arm moves, allowing reproducible placement and loading from one gel to the next. In addition, the E-Holder™ Platform allows you to load samples on one E-Gel® 96 or E-Gel® 48 gel while other gels are running in the Mother or Daughter E-Base™ combination. E-Editor™ 2.0 software for easy analysis of E-Gel® 48 and 96 gels Analysis of E-Gel® 48 and 96 gel results is fast and easy using E-Editor™ 2.0 software.‡ E-Editor™ 2.0 software takes the digital image of your gel and reconfigures the lanes into one regrouped and aligned image (Figure 19) for easy comparison, documentation, and reference with the original plate coordinates and for quick identification of important results. E-Editor™ software is available FREE of charge with the purchase of E-Gel® 48 and 96 gels and related equipment. ‡ This software is covered by a software license agreement and may be downloaded in PC format from the Invitrogen website at www.invitrogen.com/egels. E-Gel ® 96 gel E-Ho ld Figure 19—Analyze E-Gel® 48 and 96 gel results using E-Editor™ 2.0 software. er™ Plat f or m Figure 18—The E-Holder™ Platform on a liquid handling robot. The E-Holder™ Platform keeps E-Gel® 48 and 96 gels stable during robotic loading of multiple gels. www.invitrogen.com Nucleic Acid Electrophoresis Safe, sensitive DNA stains SYBR® Safe DNA Gel Stain—a better, safer stain SYBR® Safe DNA Gel Stain is a nontoxic, nonmutagenic DNA stain that can be used like ethidium bromide, either in the gel during electrophoresis or as a postrun stain. There is no need to destain before viewing and documenting your results. SYBR® Safe DNA Gel Stain is available as a ready-to-use solution, or as a 10,000X concentrate. DNA stained with SYBR® A Safe DNA Gel Stain can be detected with a visible-light transilluminator such as the Safe Imager™ Blue-Light Transilluminator, a standard UV transilluminator, or a laser-based scanner. For a complete safety solution and optimal cloning efficiency, use SYBR® Safe DNA Gel Stain with the Safe Imager™ Blue-Light Transilluminator. SYBR® Safe DNA Gel Stain displays no acute oral toxicity and greatly reduced genotoxicity, offers nonhazardous waste status B and Clean Water Act compliance, reduces waste storage and disposal costs, and provides uncompromised detection sensitivity as compared to ethidium bromide. SYBR® Gold stain outperforms ethidium bromide SYBR® Gold Nucleic Acid Gel Stain outperforms ethidium bromide and other gel stains in any gel system (Figure 20), including agarose, polyacrylamide, native, formaldehyde, glyoxal, and urea gels. Stained dsDNA, ssDNA, and RNA can be viewed with a standard 300 nm UV transilluminator, enabling you to obtain high sensitivity without using expensive laser scanners. SYBR® Gold staining can also be viewed using the Safe Imager™ Blue- Figure 20—Clear E-Gel® stained with SYBR® Gold. Identical two-fold dilutions of glyoxalated E. coli 16S and 23S ribosomal RNA were separated on 1% agarose mini gels using standard methods and stained for 30 minutes with SYBR® Gold Stain in TBE buffer (A) or 0.5 µg/ml ethidium bromide in 0.1 M ammonium acetate (B). Both gels were subjected to 300 nm transillumination and photographed using Polaroid 667 black-and-white print film and a SYBR® photographic filter (A) or an ethidium bromide photographic filter (B). Light Transilluminator. (Please note that SYBR® Gold stain does not offer the added safety benefits of SYBR® Safe stain.) Safe Imager™ Blue-Light Transilluminator eliminates UV hazards Use the Safe Imager™ Blue-Light Transilluminator to view stained gels on the laboratory benchtop (Figure 21). Light from the LED source inside the transilluminator passes through a blue filter, producing a single-intensity signal that is optimal for the excitation of SYBR® DNA-binding dyes as well as many Invitrogen protein gel stains (such as SYPRO® Ruby and Coomassie Fluor™ Orange stains). Sensitivity is comparable to that obtained with a standard UV transilluminator. Blue-light transillumination also results in dramatically increased cloning efficiencies compared to UV transillumination, and poses no danger to the end user. 10 Figure 21—The Safe Imager™ Blue-Light Transilluminator. Easily view stained gels with the Safe Imager™ Blue-Light Transilluminator. The Safe Imager™ Blue-Light Transilluminator produces brighter light and more uniform emission than conventional blue-light sources and does not damage your skin or eyes. Convenient nucleic acid markers Invitrogen offers a wide selection of products for estimation of molecular weight and mass of both DNA and RNA (Figures 22 and 23, Tables 3–5). Invitrogen’s TrackIt™ DNA Ladders are ready to load and contain two tracking dyes that indicate when maximum resolution of the samples has been achieved. The tracking dyes run outside the limits of most DNA samples and are therefore unlikely to mask a band of interest. Invitrogen’s DNA markers work well with all agarose gels, including E-Gels. TrackIt™ 1 Kb Plus DNA Ladder Cat. no. 10488-085 -bp E-Gel® Low Range Quantitative DNA Ladder Cat. no. 12373-031 bp 2,000 --bp 2,072 1,500 E-Gel® High Range DNA Ladder Cat. no. 12352-019 12,000 800 40 400 20 200 10 800 1,000 850 100 5 400 500 300 4,000 TrackIt™ 50 bp DNA Ladder Cat. no. 10488-043 10 µl run on an E-Gel® 0.8% double-comb gel TrackIt™ 25 bp DNA Ladder Cat. no. 10488-022 -bp --bp 500 TrackIt™ 10 bp DNA Ladder Cat. no. 10488-019 -bp 350 300 250 50 High DNA Mass Ladder Cat. no. 10496-016 -bp 100 4,000 50 1,200 800 400 3,000 200 10 50 1,000 4% E-Gel® 0.5 µg/lane -bp 6,000 2,000 25 100 Low DNA Mass Ladder Cat. no. 10068-013 2,000 150 2% E-Gel® 0.3 µg/lane 100 2% E-Gel® 0.3 µg/lane 10,000 75 200 200 330 200 125 100 300 0.8% E-Gel® 0.3 µg/lane 800 500 400 2,000 1,650 2,000 10 µl run on an E-Gel® 2% double-comb gel 600 500 6,000 5,000 4,000 3,000 bp 10,000 ng 100 TrackIt™ 100 bp DNA Ladder Cat. no. 10488-058 4% E-Gel® 0.5 µg/lane 0.8% E-Gel® 0.15 µg/lane 100 2% E-Gel® 0.25 µg/lane Figure 22—DNA markers for E-Gels. 11 www.invitrogen.com Nucleic Acid Electrophoresis Table 3—DNA marker selection guide for agarose gels. Agarose concentration 0.8% 1.2% 2% 4% Suggested markers* Cat. no. TrackIt™ 1 Kb Plus DNA Ladder 10488-085 500 bp DNA Ladder 10594-018 High DNA Mass Ladder 10496-016 Low DNA Mass Ladder 10068-013 TrackIt™ 100 bp DNA Ladder 10488-058 Product 10488-072 0.1–2 Kb RNA Ladder 75 µg 15623-100 TrackIt™ 1 Kb Plus DNA Ladder 10488-085 0.5–10 Kb RNA Ladder 75 µg 15623-200 High DNA Mass Ladder 10496-016 TrackIt™ 25 bp DNA Ladder 10488-022 TrackIt™ 50 bp DNA Ladder 10488-043 TrackIt™ 100 bp DNA Ladder 10488-058 TrackIt™ 1 Kb Plus DNA Ladder 10488-085 TrackIt™ λ DNA/Hind III fragments TrackIt™ ФX174 RF DNA/Hae III fragments Low DNA Mass Ladder TrackIt™ 10 bp DNA Ladder TrackIt™ 25 bp DNA Ladder TrackIt™ 50 bp DNA Ladder 10488-064 TrackIt™ 1 Kb DNA Ladder 10488-037 10068-013 10488-019 10488-022 10488-043 * Other markers may also be suitable for use. For a complete listing of our nucleic acid markers, please visit www.invitrogen.com/namarkers. Table 5—RNA Ladders for sizing single-stranded RNA in glyoxal or formaldehyde agarose gels. Quantity A Cat. no. B 0.1–2 Kb RNA Ladder 0.5–10 Kb RNA Ladder kb kb 2 10 8 1.5 6 1 4 0.8 3 0.6 0.5 0.4 2 1.5 0.3 Table 4—DNA marker selection guide for E-Gel® 48 and E-Gel® 96 gels. Agarose concentration 0.2 Gel type E-Gel® 48 1% E-Gel® 96 E-Gel® 48 2% E-Gel® 96 4% E-Gel® 48 Suggested markers * E-Gel® High Range DNA Ladde E-Gel® High Range DNA Ladder E-Gel® Low Range Quantitative DNA Ladder E-Gel® Low Range Quantitative DNA Ladder E-Gel® Low Range Quantitative DNA Ladder Cat. no. 0.1 0.5 12352-019 12352-019 12373-031 12373-031 12373-031 * O ther markers may also be suitable for use. For a complete listing of nucleic acid markers, please visit www.invitrogen.com/namarkers. 12 1 Figure 23—RNA Ladders. A. 0.1–2 Kb RNA Ladder is suitable for sizing singlestranded RNA in glyoxal or formaldehyde agarose gels. The ladder consists of ten bands in regular increments (0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.8, 1.0, 1.5, and 2.0 kb) to simplify size estimation. Ladder is supplied at 1 µg/µl in 10 mM HEPES (pH 7.2), 2 mM EDTA. B. 0.5–10 Kb RNA Ladder is suitable for sizing single-stranded RNA in glyoxal or formaldehyde agarose gels. The ladder consists of nine bands in regular increments (0.5, 1, 1.5, 2, 3, 4, 6, 8, and 10 kb) to simplify size estimation. The 1.5 kb band is more intense than the other bands, allowing easy orientation and identification of RNA bands on the gel. The RNA molecules in both ladders are polyA-tailed (40 nt) for convenient use as control templates for cDNA synthesis. Highest purity and reliability with UltraPure™ reagents UltraPure™ 10X TAE Buffer UltraPure™ 10X TAE Buffer is a sterile-filtered solution of 0.4 M Trisacetate and 0.01 M EDTA. TAE buffer is commonly used for aga- For superior results every time, trust Invitrogen’s UltraPure™ rose DNA electrophoresis. It is supplied in a 1 L plastic bottle or in reagents. All UltraPure™ reagents are made from the purest bio- a 4 L or 10 L stackable Cubitainer® Box.* A 1X TAE buffer solution chemicals for maximum reliability and superior performance. contains 40 mM Tris-acetate and 1 mM EDTA, pH 8.3. UltraPure™ Agarose * Cubitainer is a registered trademark of Hedwin Corporation, Inc. UltraPure™ Agarose resolves DNA and RNA fragments from 100 bp to >30 kb. For high resolution of small DNA, RNA, and PCR frag- UltraPure™ 10X TBE Buffer ments <1,000 bp, use UltraPure™ Agarose 1000. If low melting UltraPure™ 10X TBE Buffer is a sterile-filtered solution of 1 M Tris, point agarose is needed, UltraPure™ Low Melting Point Agarose, 0.9 M boric acid, and 0.01 M EDTA used to prepare 1X buffer for which melts at ~65°C, is recommended for resolution of DNA and polyacrylamide and agarose gel electrophoresis. This product is RNA fragments >1,000 bp. available in a 1 L plastic bottle or in a 10 L Cubitainer® Box. UltraPure™ Agarose and SYBR® Safe DNA Gel Stain Combo UltraPure™ Tris UltraPure™ Agarose and SYBR® Safe DNA Gel Stain Combos are used in buffers because of its buffering range (pH 7.0–9.0) and offered in two sizes sufficient for approximately 80 or 400 mini- compatibility with many enzymes, including restriction endo- gels. Enjoy the superior quality of UltraPure™ Agarose and the nucleases and DNA-modifying enzymes. UltraPure™ Tris [tris(hydroxymethyl)aminomethane] is widely sensitivity and safety benefits of SYBR® Safe DNA Gel Stain while saving money compared to buying these same products sepa- UltraPure™ Salmon Sperm DNA rately. The SYBR® Safe stain included in these combos is a ready- UltraPure™ Salmon Sperm DNA is a ready-to-use solution of to-use solution in 1X TAE. sheared DNA that can be used directly in the preparation of prehybridization and hybridization solutions. The DNA solution is UltraPure™ DNase/RNase-Free Distilled Water prepared from highly pure, phenol/chloroform–extracted DNA UltraPure™ DNase/RNase-Free Distilled Water is designed for use and DNase-free and RNase-free (DEPC-treated) distilled, deion- in all molecular biology applications. It is 0.1 µm membrane– ized water and is sheared to an average size of ≤2,000 bp. The filtered and tested for DNase and RNase activity. concentration is adjusted to 10 mg/ml. 13 www.invitrogen.com Nucleic Acid Electrophoresis For top-notch performance every time, look to Invitrogen for your electrophoresis needs. Order today. Ordering information Product Quantity Cat. no. 0.8% General Purpose Starter Pak (containing ethidium bromide) 1 kit* G6000-08 1.2% General Purpose Starter Pak (containing SYBR® Safe DNA Gel Stain) 1 kit* G6206-01 1.2% General Purpose Starter Pak (containing ethidium bromide) 1 kit* G6000-01 2% General Purpose Starter Pak (containing SYBR® Safe DNA Gel Stain) 1 kit* G6206-02 2% General Purpose Starter Pak (containing ethidium bromide) 1 kit* G6000-02 4% High Resolution Starter Pak (containing ethidium bromide) 1 kit* G6000-04 0.8% General Purpose 18-Pak (containing ethidium bromide) 18 gels G5018-08 1.2% General Purpose 18-Pak (containing SYBR® Safe DNA Gel Stain) 18 gels G5218-01 1.2% General Purpose 18-Pak (containing ethidium bromide) 18 gels G5018-01 1.2% General Purpose Clear E-Gel® 18-Pak (without DNA stain) 18 gels G5518-01 2% General Purpose 18-Pak (containing SYBR® Safe DNA Gel Stain) 18 gels G5218-02 2% General Purpose 18-Pak (containing ethidium bromide) 18 gels G5018-02 4% High Resolution 18-Pak (containing ethidium bromide) 18 gels G5018-04 0.8% General Purpose 18-Pak (containing ethidium bromide) 18 gels G6018-08 2% General Purpose 18-Pak (containing ethidium bromide) 18 gels G6018-02 1 kit † G6400ST 18 gels G6618-08 8 gels G8008-01 Single-comb E-Gels Double-comb E-Gels E-Gel® CloneWell gels E-Gel® CloneWell 0.8% SYBR® Safe Gels and iBase™ Starter Kit † E-Gel® CloneWell 0.8% SYBR® Safe Gels E-Gel® 48 Gels 1% General Purpose 8-Pak (containing ethidium bromide) 2% General Purpose 8-Pak (containing ethidium bromide) 8 gels G8008-02 4% General Purpose 8-Pak (containing ethidium bromide) 8 gels G8008-04 1% General Purpose 8-Pak (containing ethidium bromide) 8 gels G7008-01 2% General Purpose 8-Pak (containing ethidium bromide) 8 gels G7008-02 E-Gel® 96 Gels E-Gel® accessories E-Gel® iBase™ Power System 1 G6400 Mother E-Base™ Integrated Power Supply 1 EB-M03 1 EB-D03 2 holders EH-03 Daughter E-Base™ Integrated Power Supply E-Holder™ Platform E-Editor™ 2.0 Software and manual 1 Free download E-Gel® Opener 1 G5300-0 * Each Starter Pak includes 6 E-Gels and one E-Gel® PowerBase™ v.4. † 14 The E-Gel® CloneWell 0.8% SYBR® Safe Gels and iBase™ Starter Kit includes 18 gels and one iBase™ device. Ordering information, continued Product Quantity Cat. no. DNA stains SYBR® Safe DNA Gel Stain in DMSO, 10,000X concentrate 400 ml S33102 SYBR® Safe DNA Gel Stain in 0.5X TBE 1 L 4 L S33100 S33101 SYBR® Safe DNA Gel Stain in 1X TAE 1 L 4 L S33111 S33112 SYBR® Safe DNA Gel Stain Starter Kit 1 kit S33110 SYBR® Safe Photographic Filter 1 S37100 500 µl S11494 1 S7569 Safe Imager™ Blue-Light Transilluminator (includes Safe Imager™ Viewing Glasses and Safe Imager™ Amber Filter) 1 S37102 Safe Imager™ Viewing Glasses 1 S37103 SYBR® Gold Nucleic Acid Gel Stain SYBR® Photographic Filter Blue-light transilluminator Nucleic acid markers TrackIt™ 1 Kb Plus DNA Ladder 1 Kb Plus DNA Ladder (1 mg/ml) TrackIt™ 1 Kb DNA Ladder 1 Kb DNA Ladder (1 mg/ml) 500 bp DNA Ladder (1 mg/ml) 100 apps 10488-085 250 µg 1 mg 10787-018 10787-026 100 apps 10488-072 250 µg 1 mg 15615-016 15615-024 100 µg 10594-018 100 apps 10488-058 50 µg 250 µg 15628-019 15628-050 TrackIt™ 50 bp DNA Ladder 100 apps 10488-043 TrackIt™ 25 bp DNA Ladder 20 apps 10488-022 TrackIt™ 100 bp DNA Ladder 100 bp DNA Ladder (1 mg/ml) TrackIt™ 10 bp DNA Ladder 20 apps 10488-019 TrackIt™ λ DNA/Hind III Fragments 100 apps 10488-064 TrackIt™ ФX174 RF DNA/Hae III Fragments 100 apps 10488-037 High DNA Mass Ladder 50 apps 10496-016 Low DNA Mass Ladder 50 apps 10068-013 E-Gel® Low Range Quantitative DNA Ladder 100 apps 12373-031 E-Gel® High Range DNA Ladder 100 apps 12352-019 0.1–2 Kb RNA Ladder 75 μg 15623-100 0.5–10 Kb RNA Ladder 75 μg 15623-200 15 www.invitrogen.com Ordering information, continued UltraPure™ reagents UltraPure™ Agarose UltraPure™ Agarose and SYBR® Safe Combo UltraPure™ Agarose 1000 UltraPure™ Low Melting Point Agarose 100 g 500 g 15510-019 15510-027 80 apps 400 apps 15510-100 15510-500 100 g 10975-035 100 g 15517-022 500 ml 10 × 500 ml 10977-015 10977-023 UltraPure™ Tris 1 kg 15504-020 UltraPure™ 10X TBE Buffer 1 L 10 L 15581-044 15581-028 UltraPure™ 10X TAE Buffer 1 L 4 L 10 L 15558-042 15558-026 15558-034 5 × 1 ml 15632-011 UltraPure™ DNase/RNase-Free Distilled Water UltraPure™ Salmon Sperm DNA www.invitrogen.com © 2006 Invitrogen Corporation. All rights reserved. These products may be covered by one or more Limited Use Label Licenses (see Invitrogen catalog or www.invitrogen.com). By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. For research use only. Not intended for any animal or human therapeutic or diagnostic use, unless otherwise stated. B-065938 1106
© Copyright 2024