- Can the RunOne be used with other buffers besides TAE?
TBE and MOPS may also be used. The RunOne running unit is constructed of polycarbonate for its temperature resistance and moderate chemical resistance.
- Should we order EP-2000 or EP-2014 for use in the USA?
EP-2000 is the proper catalog number for the US version of the RunOne.
- Can the RunOne system be used for SSCP?
Yes, there are a few customers who have used the RunOne for SSCP by placing their units in the cold room and using a polyacrylamide gel cast with the RunOne casting system. The RunOne system does not provide the most flexibility in doing SSCP. It is great for initial screening, and routine analysis where the mobility shift is clearly manifested at 4 °C.
- What kind of agency approval exists for the RunOne?
The RunOne has the CE, CSA (Canadian Standard Association), and the NRTL/C marks. With these marks the RunOne system fulfills requirements for European and Canadian sales.
- What is the difference between the old RunOne and the new RunOne and how do I know what I have?
The EP-2000 was introduced August 1999 and the first generation RunOne, the EP-1000, was introduced in May 1997. Visually the two RunOne systems (catalog# EP-1000 and EP-2000) are quite similar. The most significant enhancement / improvement introduced in the EP-2000 is that the lid can now be removed without having to remove the power supply for quick access to the gel and the running chamber. In the case of the first generation RunOne, it features a mechanical interlock design, the lid is the electric connector between the power supply and the running tank. In the case of the second generation RunOne system,the lid acts as the lever for the dual waterproof on/off switches. The easiest way to tell what version of the RunOne you have is to examine whether there are two metal pins on the lid. The EP-1000 has the metal pins and the EP-2000 does not.
- How do you recommend cleaning for the RunOne?
Simply rinse out the inside of the running unit and flip it over and let air dry. We do not recommend submerging the unit in water, There is also no need to get down into the electrode area and clean the gap or groove. DO NOT WIPE OR HANDLE THE PLATINUM WIRE.
- Can I re-use my running buffer with the RunOne?
No, we do not recommend that you reuse your running buffer. This is because as the gels run, evaporation occurs. This causes the buffer concentration to change.
- Can I cast thinner polyacrylamide gels (less than 3 mm)?
No, not with the
current gel tray and cover configuration. The distance between the polyacryla-mide gel tray and cover is 3 mm.
Oxygen inhibits polymerization of acrylamide; therefore, the gel solution must completely fill the space in
between the polyacrylamide gel tray and cover in order to eliminate contact with oxygen and ensure good
polymerization.
- Is it a problem if some of the gel solution gets under the gel tray while
casting?
No. A
small amount of gel solution will flow underneath the tray, but this will not affect gel performance. After the
gel is polymerized, remove the thin layer with a tissue before running.
- When casting polyacrylamide gels, air
bubbles are sometimes trapped between the cover and the gel solution when I lower the cover onto the gel
solution. What would solve this?
Use the spatula provided to slowly lower the cover at an angle onto the gel
solution as shown in Figure 2, page 4 in the RunOne Instruction Manual.
- The gels (especially higher percentage polyacrylamide gels) are
sometimes difficult to remove from the casting stand. What do you suggest?
The small amount of gel solution
that gets under the gel tray will slightly adhere it to the casting stand. Remove the gel tray by pushing inward
on the two side tabs of the gel tray (toward the center of the gel), then pull the gel tray up and out of the
stand.
- Sometimes when removing a polyacrylamide gel tray and cover from the casting stand, air bubbles get
trapped in between the gel and the tray or in between the gel and the tray cover. Will this affect gel
performance?
No. Air pockets may be introduced when the gel tray is removed from the casting stand, but they
will not affect the running of the gel.
- Sometimes the gel tray floats above the running platform after I add
running buffer to the RunOne Cell tank. How do I get the gel tray to seat properly?
To anchor the gel to the
running platform, push it down and slide it slightly sideways.
- Why do I occasionally have trouble loading my
sample into a well?
One possibility is insufficient sample buffer density.
- The instruction said the RunOne comes with gel tray for casting polyacrylamide gels, where are the gel trays?
We found that most people only use agarose trays so in order to be environmentally friendly, we only ship the acrylamide gel trays upon request. Simply download a request form and send back to us and we'll get your trays out right away.
- What volume agarose solutions should I be using to cast gels?
When using the RunOne Casting system, use 30 ml for the wide gels and 15 ml for the mini gels. When casting a gel using a MultiCaster, use 80 ml for the long gels (blue and aqua MultiCaster) and 60 ml for the medium gels (orange and yellow MultiCaster)
- What is the maximum sample volume I can load with the RunOne?
When using any of the RunOne casting stands, you can load up to 30 µl per well on one side of the comb and 11 ul on the other side.
- Do you sell replacement agarose gel trays?
Yes, all trays are sold in packs of 5 (catalog # EP-1011 for landscape, EP-1043 for mini trays).
- The gel trays are warping after a few times of pouring agarose gels?
If the gel trays are clear, then you are using the incorrect trays, use the gray tinted trays. The clear trays are made for acrylamide gels and therefore are not very heat resistant.
Solution: If you are using the agarose trays, which are tinted gray, pour agarose at a temperature that is lower than 75 °C.
- The Power supply is blinking and there does not seem to be any current.
The number one cause of this problem is the buffer concentration. The power supply is equipped with a current limit, which monitors the current level for you. If the current gets too high, the power supply will automatically shut itself down and notify you with a blinking red light. The current limit for the RunOne power supply is set to 280 mA. This is to prevent any damage to the power supply and also to your gel.
Solutions:
- Replace buffer with fresh buffer and then continue with the electrophoresis.
- Make new buffer according to the recipe in the RunOne instruction manual. You can download our 10X TAE or 5X TBE recipe.
- Get a sample of our TAE or TBE running buffer concentrate by calling us toll free at 1-800-255-1777 or send us a sample request
- Buy your buffers from us. We offer 10X TAE in 4 1 Liter bottles (EC-1016) as well as 5X TBE also in 4 1 Liter bottles (EC-1017)
- Why is electrophoresis taking
much longer than the expected run time?
Check to make sure the running buffer was correctly prepared and
diluted to 1X concentration. Excessive salt concentration in the buffer produces higher current levels and
results in a lower voltage gradient and longer run times.
- The unit is running hot, the gel is melting
The buffer concentration is incorrect. The amount of current in the system is related to the heat generated, and the amount of current is proportional to the amount of salt. Most likely the buffer concentration is much more than 1X. For example, the temperatures of the buffers in the RunOne after a 30 minute run using a 1X TBE is 32.6°C, 90.7°F and for a 2X TBE it is 44.4°C, 111.7°F.
Solutions: use answers in 1 above.
- The unit is not running and the light on the power supply is not
blinking.
Solutions:
- There is a small green LED at the lower right hand corner of the RunOne power supply, it should be lit if a functioning power supply is plugged into the power outlet. If the green LED light is not on when the power supply is plugged in then the power supply is burnt out. Call Embi Tec to obtain a Return Authorization number and send the power supply back.
- Make sure the lid is lowered fully and firmly in place.
- The unit is leaking there is a puddle of liquid on the bench where the unit
sits.
If your RunOne lid has pins, examine the inside wall of the running tank next to the power supply and see if there are any hairline cracks. If there are hairline cracks, call Embi Tec for a Return Authorization number and send the unit back for replacement.
Condensation and then siphoning might have created a drain path from the running tank to the bottom of the unit. Empty out the buffer and let the running unit dry out.
- How can I check the conductivity of my buffer to make sure it is good?
If you have a conductivity meter or a conductivity feature on your pH meter, you can use this to measure the conductivity of your buffer. 1X TAE should give a conductance measurement of 1.25 at 19.99 mS/cm, a 1X TBE conductance measurement is 0.87 at 19.99 mS/cm.
Another way to get an idea of the relative conductivity of your buffer is to use the ending temperature of a 30-minute run. Here are two charts of different concentrations of TAE and TBE and their respective ending temperature after a 30-minute run. We have provided this chart for both Fahrenheit and Celcius scales.
- What kind of gels do you make?
Horizontal Agarose and Polyacrylamide gels for DNA and RNA analysis.
- What percentages of agarose gels do you make?
1, 2, 3, 4%
- Do you make gels with Ethidium Bromide?
Our agarose gels are available with EtBr stain, the polyacrylamide gels
are not.
- What buffer types do you make?
TAE and TBE
- Does your precast gel only run in your unit?
No, run it in any unit with a large enough platform. Of course you will get a perfect fit in our unit.
- How stable are your precast gels?
6 months at room temperature
- How much Ethidium Bromide do you use in your gels?
0.75 µg/ml
- What is the separation range of your precast gels?
Depending on the percentage and gel type you choose, you can separate fragments from 20 bp up to 10,000 bp
- How many different size gels do you make and why?
Currently we have 6 different sizes of gels. Take a look through and you're sure to find a gel will fit your application and box.
- Are your precast agarose gels compatible with multi channel pipette loading?
Yes, with a few exceptions, every one of our gels was designed with multi channel loading in mind.
- Do you have a gel that can run all 96 samples of a PCR plate at a time?
Yes, we have 3 different gel sizes and a variety of comb configurations to accommodate 96 samples at a time. We even have gels, which are formatted exactly like your PCR plate so you don't have to interpret your gels. Best of all, we add additional wells to each row so you don't have to sacrifice a sample for your standards.
- Do you use molecular biology grade agarose?
Yes we do.
- Do you use NuSieve agarose for your 4% gels?
No.
- Does the long gels fit into the RunOne, it looks so large?
Amazingly, it does! All of our gels, except the very largest (Excel), were designed to fit into the RunOne unit.
- What volume can I load into your gels?
Depending on the gel you pick, you can load anywhere from 10 - 30 µl. We also have preparative gels that can hold up to 112 µl
- How are your gels shipped?
Most orders are shipped the same day or next day from the purchase order. To ensure their quality when they arrive to you, all gels are shipped 2 days or less.
- What kind of gels do you make?
Horizontal Agarose and Polyacrylamide gels for DNA and RNA analysis.
- What DNA standard is appropriate for the PAG gels I am using?
We recommend using a 100 bp and /or 20 bp standard on our horizontal polyacrylamide gels. The 100 bp and 20 bp standard can also be mixed 1:1 and used as a single standard on one lane of gels to give a finer molecular weight ruler when estimating molecular weight.
- Do you make gels with Ethidium Bromide?
Our agarose gels are available with EtBr stain, the polyacrylamide gels
are not.
- What buffer types do you make?
TAE and TBE
- Does your precast gel only run in your unit?
No, run it in any unit with a large enough platform. Of course you will get a perfect fit in our unit.
- How stable are your precast gels?
6 months at room temperature
- What is the separation range of your precast gels?
Depending on the percentage and gel type you choose, you can separate fragments from 20 bp up to 10,000 bp
- How many different size gels do you make and why?
Currently we have 6 different sizes of gels. Take a look through and you're sure to find a gel will fit your application and box.
- I have been a customer for your precast agarose gels and I tried your polyacrylamide gels to separate some of our smaller PCR products but got no bands, what is happening?
Solution: Stain the polyacrylamide gels as they do not contain ethidium bromide. Remove the gel from the gel tray and cover before staining.
- How long should I stain my polyacrylamide gel?
Stain the gel in 1 µg/ml of ethidium bromide for 15 to 30 minutes at room temperature. Destaining is not necessary. Typically, you can visualize your gel and see the bands after 5 minutes of staining.
- How are your gels shipped?
Most orders are shipped the same day or next day from the purchase order. To ensure their quality when they arrive to you, all gels are shipped 2 days or less.
- What is MOPS gel?
RunOne MOPS gels are 1.25% agarose gels in MOPS (3-N-morpholinopropanesulphonic acid) buffer, pH 7.0. The neutral pH feature of the MOPS gels makes them ideal for RNA analysis as RNA is base labile.
- I am going to check RNA for integrity, do you have a gel for this
application?
Our MOPS gel kit comes complete with the precast MOPS gel, running buffer and denaturing sample buffer.
- The RNA sample buffer does not come with any instruction, what is the proper
procedure?
Solution:
The RNA denaturing sample buffer is 2X. Dilute your sample with the sample buffer on a 1:1 ratio. Mix well and then heat at 15 minutes at 65 °C. Cool immediately and sample is ready to be loaded.
- Does your MOPS gel contains formaldehyde?
No, it does not. All of the denaturing chemicals are in the sample buffer. Traditionally, MOPS gels or gels for resolving RNA to do Northerns are run overnight with a denaturant such as formaldehyde in the gel. These gels are typically much larger for higher resolution and larger sample load. Run time for the RunOne MOPS gel is under 2 hours, during this short time the heat denaturing step and the denaturants in the sample buffer will prevent your samples from renaturing.
- I am using your MOPS gel but I am not getting any bands, just a smear.
The most likely explanation is the RNA is not totally denatured.
Solution: Use a denaturing sample buffer like our RNA Denaturing Sample Buffer (Catalog number EC-1021) and denature per protocol.
- How are your gels shipped?
Most orders are shipped the same day or next day from the purchase order. To ensure their quality when they arrive to you, all gels are shipped 2 days or less.
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