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View additional product information for BenchPro™ 2100 Plasmid Processing Station - FAQs (MC1001)
35 product FAQs found
It is completely normal for this to happen during the run and we often see it, but haven't had it lead to any problems in the instrument. You can try widening the holes with the piercing tool, which will help any of the liquid run back into the tray. More debris on the top of the foil can occur when there is too much of a high OD culture added into the system; therefore, check this and scale back accordingly. After heavy use of the instrument, you may see some of this debris ending up in the drawer and occasionally on the tracks of the instrument, but it won't hurt the instrument. However, if it's a concern, we would recommend taking some Kimwipes wipers, moistening them with 70% ethanol, and wiping it down.
Please follow the sequence of steps below to try to release the drawer lock:
- Power-off the instrument.
- Remove card (if already inserted).
- Press the drawer inward (holding the back of unit) to ensure the lock mechanism is not catching, then release pressure.
- Power-on the instrument.
- Press the drawer inward (holding the back of unit) again and attempt to open door.
- The software sends an unlock signal to the door lock solenoid during the start-up process. If after a few attempts of steps 1 through 5 the door still does not open, please call our Technical Support team.
This is usually caused by a sudden dip in the air flow (an air system problem, usually not related to the instrument itself). During the BenchPro 2100 procedure, the lysis step and elution step are the two steps that require higher air pressure, so this error message is usually seen at these two steps. If this error happens during the lysis step, we recommend restarting from the beginning, with a new culture, new card, and new reagent tray. There is no way to recover the sample.
If this happens at the elution step, you can see if you can get any eluted DNA. Hopefully some DNA has already eluted out (usually most DNA is eluted out in the first 1-2 minutes of the elution step). If this happens right before the elution step, with no solution in the elution tube yet, you can empty out the reagent tray reservoirs and refill them with 1.5 ml of TE or water in the right reservoir, then set up the run with the same tray and card and run the protocol again.
These are valve errors. The error number tells which valve is failing.
Typically, “valve=1/Error Code 52=1” means an air supply issue, as valve 1 often fails if the air pressure or flow is on the low side. If you see this error message, please measure the air pressure and flow rate. Once verified, cycle the power and restart the machine. It should pass the self-check test. However, if the problem persists even if the air pressure and flow rate meet the instrument's needs, it means the instrument needs to be serviced due to liquid or debris in the air lines. The most common cause for this failure is using too-rich medium or too many cells.
For “valve=5/Error Code 52=5” or any number except 1, try to reboot the instrument. If the same valve error shows up, it usually suggests that this specific valve is failing. If after rebooting, you see a different error number, please check to see if the air pressure and flow rate meet the requirements.
There are three possible reasons for this:
- The variability of the air flow will reduce the amount of plasmid DNA recovered from the precipitator membrane. An expected elution volume recovery should be between 800 and 1,000 µl.
- If larger plasmids are in use, the DNA size may impede the precipitator membrane enough to reduce the final elution flow. We have verified that elution volume (and yield) may be reduced if plasmids are large or exceed the recommended range.
- If you using enriched medium (TB, Circlegrow, etc.), the low elution volume may be due to overloading of the system, causing the pump to overload during the first few pumping cycles, causing blow-back into the reagent tray and blowing out some of the elution volume. A drop in house air pressure can also cause a low elution volume.
Yield can be variable, but should be between 500 µg and 1 µg per purification. This range is a result of: air pressure and air flow into the instrument, cell amounts (due to volume, strain, or type of medium), culture growth phase, plasmid size, and the ori on the plasmid.
The two most common seen causes for low/no yield are: low air pressure and/or flow rate (the instrument requires air flow rate of 4 cfm at 90 psi) and overloading (we recommend using 100-125 mL of LB culture at OD 2.0).
No, the current version of your software will not allow you to interrupt and subsequently continue your run.
No, the current version of the BenchPro 2100 cannot be connected to any laboratory information management systems.
Yes. You can resuspend the pellet in LB media or sterile water such that the OD at 600 nm is 2.0.
No. Purification of BAC clones is not optimal due to their large size. The BenchPro 2100 instrument is capable of purifying plasmids from 2.7 to 20 kb.
We have tested DH10B-T1R, DH5alpha, JM109, TOP10, and XL1-Blue in-house, and expect that any strains derived from these will also work.
Note: endA+ strains have been shown to work, but there may be nuclease activity in the final purified plasmid.
For best results, follow these guidelines:
- Use a bacterial culture at a cell density of approximately 10E9 cells/mL, or an optical density of 2.0 to 2.4 at 600 nm (OD600). Use bacterial culture in transition between exponential and stationary phase.
- Use a high copy number plasmid to obtain a good yield of plasmid DNA. High copy number plasmids typically yield 2-6 µg DNA/mL LB culture grown overnight.
You can clean the surface of the BenchPro 2100 instrument with a damp cloth. Do not use harsh reagents or solvents to clean the unit. Clean the bottom of the instrument drawer with a light spray of 70% ethanol and then wipe off the ethanol with a paper towel.
The purified plasmid is ultrapure and suitable for all downstream applications, including those requiring the highest purity, such as transfection of mammalian cells, automated and manual DNA sequencing, PCR amplification, in vitro transcription, bacterial cell transformation, cloning, and labeling.
On average, you can process 0.7 to 0.9 g of wet pellet biomass on this instrument. However, this is subject to variability when working with different strains.
Yes. Resuspend the cell pellets in 100 mL of LB medium before you load the mixture onto the cell liner tray.
The instrument is capable of purifying plasmids from 2.7 to 20 kb.
The BenchPro 2100 instrument needs a manual setup time of less than 5 minutes. After setup, the instrument takes about 90 minutes for the entire purification process. You do not need to be present during the purification process.
We offer an Air Compressor (Cat. No. MC5001), which is validated for use with the BenchPro 2100 Plasmid Processing Station.
Air compressors from other vendors could be used too, if they meet the following requirements:
- Regulate pressure output between 90 and 120 psi
- Regulate pressure output flow at greater than 4 cfm, when measured at 90 psi
- Output clean and dry air constantly
Always store the BenchPro 2100 Plasmid Card and BenchPro 2100 Reagent Tray in the supplied box to prevent any damage to the card sippers or to the tray foil. Store both at room temperature
No. The cell liner tray and lid are not reusable. They should be disposed of after each procedure. Each BenchPro 2100 reagent kit comes with a supply of cell liner trays and lids. Cell liner trays are not currently available for standalone purchase.
The waste tray is reusable and is made of material that can be autoclaved. You can also order new waste trays.
No, the BenchPro 2100 Plasmid Purification Card is not reusable. During the purification, E. coli cells are captured and lysed within the card, making subsequent reuse impossible.
You can run one or two samples simultaneously on the BenchPro 2100 Plasmid Processing Station
No, the instrument works using positive air pressure, and cannot work on vacuum. It works using house air, which is the orange line in the lab that forces air out.
You will need an in-house air supply outlet (90-120 psi, > 4 cfm, constant flow rate), or a compressor unit with the same specifications, that can be connected to the BenchPro 2100 Plasmid Processing Station.
You can use a Hedland air gauge to measure the air flow rate for the BenchPro 2100 Plasmid Purification instrument.
A self test is performed each time a run is started and takes approximately 6 minutes to complete.
Yes, you can replace TE with water. Carefully remove all TE solution from the TE-containing well (the one closest to the elution tube slot on the reagent tray), rinse the well twice with purified water and then fill with 1.5 mL purified water. You can also pipette out up to 700 µL of TE, so that the final DNA concentration could be higher. We would not recommend using less than 750 µL TE buffer.
We recommend to use 100-125 mL of LB culture at about OD 2.0. We don't recommend using enriched media like TB, YT, etc., as a high cell density will clog the membrane and decrease DNA yield from the purification. If you already have culture in TB, dilute 50 mL culture wtih 50 mL media.
While yield and purity will differ depending on the size of your vector and copy number as well as cell strain used, a high copy vector (pcDNA3.1) has been tested in several popular strains including C600, DH10B/T1R, DH5alpha, JM109, TOP10, and XL1-Blu, demonstrating an average yield above 500 µg. Purity-wise, isolated DNA usually have both A260/A280 and A260/A230 ratios in the range of 1.8-2.0. Endotoxin is usually < 10 EU/µg (3 EU/µg average).
Yes, we offer the BenchPro 2100 Plasmid Purification System, which is a fully automated maxiprep purification system.
We recommend only running one BenchPro 2100 Maxi Plasmid Processing Station instrument for each air compressor.
The minimum specification for the BenchPro 2100 Maxi Plasmid Processing Station air flow rate is 3 CFM. There is no maximum air flow rate specified.
There are 5.58mL of isopropanol and 5.33mL of 70% ethanol per tray in the BenchPro 2100 Plasmid Purification Card and Reagent Tray Kit.