Troubleshooting and FAQ
To anchor the gel with the gel tray to the running platform, push it down and slide it slightly sideways.
Check to make sure the running buffer was correctly prepared and diluted to 1X concentration (or to 0.5X concentration if running a gel prepared with 0.5X buffer). Excessive salt concentration in the buffer produces higher current levels and results in lower voltage gradient and thus a longer run time.
If the green LED does not light, check the fit between the lid, running tank and power supply. The Lid should fit flush with the running tank and the power supply should slide firmly into place. Also check the power source or the wall outlet. If all components are in place and the green LED still does not light, call Embi Tec at 1-858-684-3190 for tech service
The current has exceeded the safety limit of 400 mA and the RunOne Power Supply has shut down. Check to make sure the running buffer was correctly prepared and diluted to 1X concentration. If the running buffer has been reused, it is time to change the buffer. We highly recommend using fresh buffer after each run. If you are reusing your buffer, we suggest replenishing lost buffer with DI water - do NOT replenish with running buffer.
No. Do not add running buffer to replenish any buffer loss. The more running buffer you add after each run, the more concentrated your running buffer becomes. Excessive salt concentration in the buffer will result to:
1. Higher current levels and lead to excessive heat generation. Eventually, the buffer temperature can be high enough to melt your gel.
2. If the current exceeds the safety limit of 400 mA, the RunOne Power Supply will shut down to prevent overheating. You risk losing your samples.
3. Excessive salt concentration in the buffer produces higher current levels and result in a lower voltage gradient and thus requires a longer time to run a normal gel.
Check to make sure the gel was properly placed on the running platform with the wells closer to the power supply. DNA migrates from cathode(-) to anode(+) in the RunOne Unit as indicated by the polarity arrow on the tank.
Simply rinse out the inside of the running tank with DI water and flip it over, propped up, to air dry. We do NOT recommend submerging the unit in water or getting down to the electrode area to clean the gap or groove.
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.
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.
TBE and MOPS may also be used. The RunOne running unit is constructed of polycarbonate for its temperature resistance and moderate chemical resistance.
EP-2000 is the proper catalog number for the US version of the RunOne.
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.
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.
Make sure that the power supply is properly plugged in and check for any loose connections.
Check the Voltage Select button. Is it responsive? If not, call Embi Tec for tech service.
No. A small amount of gel solution will flow underneath the tray; this will not affect gel performance. After the gel solidifies, just remove the thin layer of gel (with tissue paper) before running the gel.
One possibility is insufficient sample buffer density. Check your sample loading buffer concentration and dilute with sample accordingly. For example, when using a 6X sample loading buffer, mis every 1 volume of loading sample buffer with 5 volumes of sample.
If the standard is stained properly, try lengthening the staining time. If the sample is still not seen, there may not be enough DNA in the sample.
When using the RunOne Casting System, use 40 ml for the Landscape gels and 20 ml for the Mini gels. When vasting a gel using a Multicaster, use 85 ml for the Long gel (Blue and Aqua MultiCasters) and 65 ml for the Medium gel (Orange, White and Yellow MultiCasters).
When using the RunOne White Comb to cast gels, you can load up to 30 µl in the wider wells and 15 µl in the smaller wells. When using the Aqua, Blue, White or Yellow MultiCaster Combs to cast gels with the different MultiCasters, you can load up to 30 µl in the wider well and 11 µl in the smaller wells.
Do NOT use glass containers when making GelGreen Gels. It is very important to use polypropylene containers instead of glass containers because glass surfaces will absorb GelGreen and render it unable to stain the DNA.
1, 2, 3, 4%
No, run it in any unit with a large enough platform. Of course you will get a perfect fit in our unit.
Yes, with a few exceptions, every one of our gels was designed with multi channel loading in mind.
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.
Depending on the percentage and gel type you choose, you can separate fragments from 20 bp up to 10,000 bp.
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.
GE-3600 is a Landscape gels so there are 10 gels. Typically Long, Medium, X-Long and X-Wide gels come 5 gels per box. Portrait and Landscape gels come 10 per box.
No, gels are already in a tray for easy handling and transport. You can run the gel while it's in the gel tray.
The tray is warping due to the high temperature of the agarose. Wait for the agarose to cool before pouring it into the gel tray.
The combs we use and sell are typically 1mm to ensure a crisp sharp band. Any other thicknesses will be specified.
For our standard Casting System, the trays are printed for better visualization of the wells. If you are using our MultiCasters, visualization plates are included.
Before pulling the comb out, pour a little bit of buffer on top near where the wells are and wiggle it a little before completely pulling the comb out.
We have RNA MOPS gels in the GE-6000 series and also RNA MOPS kits that are complete with MOPS running and denaturing buffer as well.
For optimum performance, we suggest using GelGreen or other fluorescent dyes similar to Sybr Green I.
Gels and dyes at or around 470nm ±30nm.
We suggest wiping it gently with some DI water and a soft cloth. In most cases, because this is a prep unit, a cutting mat is put underneath the gel so the surface itself shouldn't even be getting too dirty!
The Sapphire is water resistant, but not water proof. We suggest wiping the bottom of the gel tray with a paper towel before transferring it from the buffer tank to the light box.
It could be that you are trying to view the gels immediately after it has come from the running tank. We suggest cooling the gels before viewing them as it reduces (if not eliminates) condensation creating a brighter signal for visualization. An alternative reason would be that there is not enough loading sample.
No. Unlike the UV light, the PrepOne’s blue light allows for its users to view the gel and access the bands and cut it with no UV exposure giving a better quality DNA.
In the USB stick there is a file called "test_in01.csv" you can use this as a template for creating the files you need.
Open it up in Excel and you should see 3 columns that have text in it.
*Column A tells you the plate name, In this case it is "plate 1" and "plate 2". This tells you that this document will generate files for 2 plates (in01.emb, in02.emb)
*Column B tells you the order in which the wells will progress in.
*Column C to Column XXX tells you the simultaneous wells.
For example, for plate 1 A1 and B2 will illuminate at the same time, press GO then C1 and C2 will illuminate at the same time. The next time you press GO, C3 will illuminate, then G10 and so on.
In the case you Column D, Row 1 is filled in with P24, Then when you first start, A1 and B2 and P24 will illuminate at the same time.
Once you are done, save the CSV file and now you have to convert it so the LightOne understands it.
Depending on whether you are using a 384-well plate or a 96-well plate, choose the appropriate converter. "CSV_Conv384" for 384-wells or "CSV_Conv96" for 96 wells.
Click on the appropriate one and the program should load without having to install anything. Once the program loads, go to File-->Open-->"test_in01.csv"
It should look like
Click on the icon with the little green arrow and the piece of paper or click on CreateEmbitecFiles
It should create files in01.emb, in02.emb, a plate list and an error list (if errors exist). Put the inXX.emb files in the Embi Tec USB stick and plug it into the LightOne.
It allows you hand-free access
Press arrows simultaneously