Documents & Support

We offer the following systems for drying of protein gels:

DryEase Mini Gel Drying System (Cat. No. NI2387): Simple-to-use kit for even, crack-free drying of mini gels that dries gels evenly, using passive evaporation. Gels dried with this system are suitable for fluorography, densitometry, autoradiography, or permanent gel storage. Each DryEase Mini-Gel Drying System contains the following components:

- 2-DryEase Mini-Gel Drying Frames (includes 2 frames + 8 clamps) (Cat. No. NI2380)
- 1-DryEase Mini-Gel Drying Base (Cat. No. NI2300)
- 200-DryEase Mini Cellophane precut sheets (Cat. No. NC2380)
- 500 mL Gel-Dry Drying Solution (Cat. No. NI2100)
Large Gel Drying Kit (Cat. No. NI2207): Provides the same even, crack-free gel drying seen with the DryEase Mini-Gel Drying System but is designed to accommodate larger gels or 4 mini-gels. A base for the system is not required.

Each Large Gel Drying Kit contains the following components:

- 1-DryEase Large Gel Drying Frame (includes 8 clamps) (Cat. No. NI2200)
- 100-DryEase Large Cellophane precut sheets (Cat. No. NC2200)
- 500 mL Gel-Dry Drying Solution (Cat. No. LC4025)

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

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Please see below for possible causes and solutions:

- Air trapped between cellophane layers. Apply adequate Gel-Drying solution to prevent bubbles between the gel and cellophane assembly.
- Tension on cellophane too strong. Take care not to stretch the cellophane during dryer assembly.
- Rough edges or small cracks on gel edges. Trim rough edges from the gel to eliminate starting points for cracks.
- Cellophane slips during drying. Make sure clamps hold all four edges of the dryer assembly firmly.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

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Gels turn white when they are dried too rapidly, usually in an environment that is too ventilated, drafty, or warm. You can remedy this problem by dabbing the gel with deionized water on a Kimwipe tissue to rehydrate the white areas. Re-dry the gel in a dark, dry, non-drafty place, such as a drawer or cabinet.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

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Usually it is not necessary to peel off the cellophane. If your signal is very faint and you are concerned about getting an exposure, you can rub the top surface of your gel with a damp towel and then peel off the cellophane. Then air dry the gel for a few minutes and perform autoradiography.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

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The pore size of DryEase cellophane is 40 microns.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

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We recommend using the Large Gel Drying Kit (Cat. No. NI2207) for drying E-PAGE gels. It is also possible to air dry E-PAGE gels. The E-PAGE 96 gels will need at least 4 days for complete air drying. Vacuum drying is not recommended - the thickness of these gels makes them especially prone to cracking.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

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Large inserts should be gel-purified prior to cloning. Gel purification of long PCR products often improves efficiency, as smaller non-specific bands that may not be visible on the gel will interfere with cloning of the insert of interest. However, the TOPO XL-2 Complete PCR Cloning kit is supplied with the PureLink Quick Gel Extraction and PCR Purification Combo Kit for added flexibility allowing for faster PCR purification when inserts are smaller, while primers and dNTPs still need to be removed.

Keep in mind that the TOPO reaction does have a strong size bias, and that any smaller fragments present in the PCR reaction would clone much more efficiently than the fragment of interest, even if the smaller fragment is not readily visible on a gel. Therefore, it is highly recommended that the PCR product of interest be gel-purified.

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Large inserts should be gel-purified prior to cloning. Gel purification of long PCR products often improves efficiency, as smaller non-specific bands that may not be visible on the gel will interfere with cloning of the insert of interest. The TOPO XL-2 Complete PCR Cloning Kit is supplied with the PureLink Quick Gel Extraction and PCR Purification Combo Kit for added flexibility allowing for faster PCR purification when inserts are smaller, while primers and dNTPs still need to be removed.

Keep in mind that the TOPO reaction does have a strong size bias, and that any smaller fragments present in the PCR reaction would clone much more efficiently than the fragment of interest, even if the smaller fragment is not readily visible on a gel. Therefore, it is highly recommended that the PCR product of interest be gel-purified.

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Here are possible causes and solutions:

- Voltage is too low: 1 mm thick polyacrylamide gels (Mini and Midi Gels) should be transferred at 20 V, E-PAGE Gels at 25 V (approximately 15 V/cm field strength).
- Power supply is inappropriate for semidry transfer: Some power supplies will shut off or blow a fuse when run at the conditions required for semi-dry transfer. Semi-dry transfer requires low voltage (20 V) and high current. Check with the manufacturer of the power supply to determine whether it is appropriate for semi-dry transfer.
- Transfer was performed for too short a time: Increase the amount of time for transfer. Typical transfer times range from 30 to 60 minutes.
- Transfer sandwich was assembled in the wrong order: The Invitrogen Semi-dry Blotter is configured with the cathode on the top, and anode on the bottom. This results in a downward transfer of proteins from the gel onto the membrane. Follow the instructions carefully when assembling the transfer sandwich.
- The pH of the transfer buffer is too close to the isoelectric point of the protein: The transfer buffers should be at the optimal pH if prepared as described in this manual. Do not adjust the pH with acid or base as this will increase the conductivity of the buffer and result in higher current during transfer.
- Too much methanol is present in the transfer buffer: Reducing the amount of methanol can help elute proteins from the gel, but can reduce binding to nitrocellulose membranes.
- High-percentage gels restrict transfer: Higher percentage acrylamide or crosslinkers can restrict elution of proteins. Use the lowest percentage acrylamide possible to separate your proteins.
- Puddles of buffer were present on the plates, allowing the current to bypass the stack: Always clean up the lower plate before closing the lid of the transfer apparatus. Do not squeeze the stack excessively, as this removes transfer buffer from the blotting paper and also creates puddles that the current can pass through.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

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The Ion 550 Single Chip Supplemental Kit contains pre-packaged templating supplies and sequencing reagent cartridges required to support eight single chip Ion Chef runs and eight sequencer initializations with the Ion 550 Kit-Chef. The Ion 550 Single Chip Supplemental Kit includes the following components:

- Ion S5 Chef Supplies (4 boxes) (ships at room temperature; store at room temperature)
- Ion S5 Sequencing Solutions (1 box) (ships at room temperature; store at room temperature)
- Ion S5 Sequencing Reagents (1box) (ships on wet ice at 2-8 degrees C; store at -20 degrees C)
IMPORTANT! Do not store the Ion S5 Sequencing Reagents (Part No. A27768) on dry ice or in a closed environment containing dry ice.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

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No, most transfer applications require high current and therefore, we recommend using a high current power supply like PowerEase Touch 350W Power Supply.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

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We recommend using the Lumio gel sample buffer that is supplied with the Lumio Green Detection kit, for in-gel detection. We do not recommend using the E-PAGE loading buffer. There is no need to remove the E-PAGE gels from the cassette to visualize Lumio fusion proteins.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

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We recommend using the following instructions for Graphite conductive adhesive, alcohol based for maximum adhesion:

1. Prepare the substrate surface: Before coating substrates, we recommend that they be clean and dry. You may use a solvent wipe.
2. Mix the adhesive: This product is supplied in its concentrated form. Please note that it is thixotropic in nature and has the tendency to gel on standing. Preparation includes thorough agitation, then dilution with isopropanol to yield the required consistency for the application of choice.
3. Apply the adhesive: Diluted graphite conductive adhesive may be applied with the use of a spray, brush, dip, or roller. We do recommend using a spray, as it yields the most uniform coverage.
4. Cure the adhesive: This product air dries within five minutes under normal temperature and humidity. Complete cure properties will develop in about two hours with consideration of film thickness and drying conditions. To achieve optimum coating qualities in a shorter curing cycle, after air drying, bake for 5 minutes at 75°C (167°F).

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In order to avoid heat-induced strand scission of RNA that occurs in the presence of divalent cations (e.g., Mg++), we have increased the EDTA content of the Formaldehyde Load Dye supplied with the NorthernMax and NorthernMax Plus Kits to a level significantly higher than the formulations found in many common laboratory guides.

When ethidium bromide is added directly to the Formaldehyde Load Dye before electrophoresis, the ethidium staining of the RNA is reduced compared to typical dye formulations. There is no reduction in ethidium bromide staining when gels containing samples run with Formaldehyde Load Dye are stained post-electrophoresis.

Another effect of the relatively high EDTA concentration in the Load Dye is that the sensitivity of the positive control reaction supplied with the kit is increased relative to typical formaldehyde loading dyes. In other words, when the Control Template RNA from the kit is electrophoresed, blotted and probed with either RNA or DNA probes produced using the control templates from the kit, a stronger signal is seen from samples run in Ambion Formaldehyde Load Dye than those run in loading dyes with less EDTA.

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