Fully Automated Plasmid Purification
Maxi, Mega, & Giga Plasmid Prep
Highly pure transfection grade plasmid DNA
Advanced silica chemistry
Highly pure, low endotoxin plasmid DNA for every scale
The PhyNexus AutoPlasmid MMG is the industry’s first and only fully-automated bench-top instrument for transfection quality plasmid purification at the maxiprep, megaprep and gigaprep scales. The automated 2 channel instrument delivers a robust and flexible solution capable of working with LB or TB media, one-touch or customizable methods, and delivering maxi, mega or giga prep scale transfection quality plasmids. The AutoPlasmid MMG allows research laboratories to increase productivity of lab specialists, improve results through automation and remove the tedium of manual plasmid preps. The AutoPlasmid MMG takes about 5-10 minutes to set up, then hit “start” and walk away until your plasmids are ready.
With Dual Flow Chromatography fluid flows back and forth through the pipette tip column. Interactions of plasmids with the column are pushed to completion. Through this fluid motion, columns are completely loaded allowing high recovery of the plasmids.
The AutoPlasmid MMG provides consistency and reliability due to the ability to purify in the presence of particulate. The process is rugged – there are no restrictions on the type of plasmid sample. 1-4 samples can be processed per run. Depending on scale, 3-12 preps can be performed every day. Recoveries of the purified plasmids are into 4 flip top vials with a bio compatible buffer. The plasmids are transient transfection quality and can be used for any desired application.
Kits are color coded and pre measured for each scale and it only takes about 5-10 minutes hands-on time to set up the instrument, pour in the prepared sample and start a run on the AutoPlasmid MMG instrument. At the end of the run, your transfection grade plasmids are deposited in vials, which are ready for use.
What are the expected yields from a Maxi, Mega or Giga prep?
The number of bacterial cells and the amount of plasmid DNA per cell can vary greatly between cultures based on many factors (strain of E. coli, plasmid copy number, media type, growth conditions). The maximum yields from a Maxi, Mega and Giga prep are 1.5 mg, 5 mg, and 10 mg, respectively. The typical yields from a Maxi, Mega and Giga prep are 750 µg, 3.5 mg, and 7 mg, respectively.
What are the advantages of the PhyNexus Maxi, Mega, and Giga prep kits?
PhyNexus plasmid purification kits offer full automation that is reliable and requires minimal maintenance. The kits are robust and compatible with a wide range of starting culture volumes. The purified plasmid DNA is transfection quality and can be eluted at high concentrations, eliminating the need for an additional ethanol precipitation step
How is the AutoPlasmid MMG different from other technologies?e
The AutoPlasmid MMG was designed to handle a wide range of starting samples.. Reliability is one of the most important qualities of laboratory equipment, and the AutoPlasmid MMG was built to be robust. Combining a simple set-up with broad sample input requirements gives the AutoPlasmid MMG a distinct advantage over conventional manual purification kits.
What other consumables or accessories are needed besides the Maxi, Mega and Giga prep kit?
Required:
Recommended:
Does the kit need to be refrigerated?
No, the kit does not have to be refrigerated. The kit comes with RNase A and it is recommended to store that at -20 °C.
What is the expiration date on the kit?
The kit is guaranteed for 1 year after manufacturing date.
Is the purification process compatible with TB and other rich media?
Yes, the PhyNexus plasmid purification columns are compatible with TB and all other rich media. Because the AutoPlasmid MMG was has a high capacity buffer system, we recommend using rich media to obtain the highest amount of plasmid DNA.
Is the plasmid purification process compatible with LB media?
Yes, the PhyNexus plasmid purification columns are compatible with LB and other simple media. However, the amount of plasmid DNA achieved using these media types is typically lower.
What size plasmids are compatible for purification on the AutoPlasmid MMG?
The AutoPlasmid MMG and kits are compatible with all plasmid types. It is recommended that low copy plasmids be cloned to medium and high copy vectors to obtain higher yields when possible.
What host cells are compatible for purification on the AutoPlasmid MMG?
Competent cells used for transformation of plasmid and ligated material can be generated in house or purchased. Sources and types of competent cells include XL1-Blue (Agilent) and DH5α (Invitrogen) for transformation efficiency of 1 x 106 transformations per microgram of DNA. These sources and laboratory preparations of competent cells are recommended when DNA is abundant (1-10 ng per reaction).
When DNA is less abundant, commercial sources of ultracompetent cells are recommended. These cells feature genes or genetic mutations that increase competence, decreases loss of DNA, resistance to and/or decreased endonuclease activity. Commercial strains of ultracompetent cells include One Shot TOP10 cells from Invitrogen and XL2-Blue from Stratagene which report transformation efficiency of 5 x 109 transformations per microgram of DNA.
Ultracompetent cells are also recommended for transformation of large clones. In addition, if processing times are limited, Mach 1 cells may be used for their increased growth efficiency. Double times are reduced from 74 minutes to 50 minutes when compared with DH5α. Plasmid preps can be carried out in as little as 4 hours after inoculation. However, the yield is dictated by the downstream requirements.
What culture volumes should I grow?
For best results, we recommend using a culture volume based on cell mass. A denser culture will result in a larger cell pellet wet weight, while a less dense culture will result in a smaller cell pellet wet weight. A low cell mass will result in lower than expected DNA yields regardless of the culture volume used. Therefore, it is always better to use denser cultures to achieve high DNA yields. Typically for maxi, mega and gigapreps, PhyNexus recommends cultures volumes of 400 mL, 1.5 L, and 3 L, respectively. If you need more plasmid DNA, you can always use more culture.
How long should I grow the culture?
Cultures should be grown to obtain large pellet wet weights. Generally, cultures should be grown for at least 16 hours.
At what RPM should I shake the culture?
The incubator setting is dependent upon the amplitude of the shaker, type of flask being used, and the culture volume to flask ration. For a New Brunswick I26 Incubator/Shaker with an orbit of 2.5 cm 350 rpm is used to grow cultures.
Is there a recommended culture flask?
It is recommended to use baffled flasks such as the Thomson Ultra Yield.
What should be used to seal the culture flasks?
AirOtop Enhanced Seals from Thomson have a very high rate of air exchange.
How to I purify more plasmids?
The flexibility and robustness of the AutoPlasmid MMG and kits mean that ideal mass of plasmid can be achieved. Generally, if more plasmid is desired, then the user can grow larger cultures.
Why is it recommended to grow larger cultures than what is typically needed for a manual prep?
Automation of a process is not simply automating a manual process. Instead, a whole new technology must be developed to find solutions for manual steps that are not feasible to automate. It is recommended to input an excess amount of plasmid in order to guarantee the desired yield is achieved. The benefit of reproducible walk-away automation outweighs the cost of growing extra E. coli culture. To get more, grow more.
What form is the starting sample, the overnight culture, or cell pellet?
The AutoPlasmid MMG procedure begins with a cell pellet generated by centrifugation of an overnight culture volume.
Does the purification process use anion exchange?
No, the plasmid purification process uses enhanced silica chemistry. This results in transfection grade plasmid DNA at usable concentrations without the need for ethanol precipitation.
Why is elution done using gravity?
Plasmid release from PhyTip columns requires gravity elution for high performance and high concentration. The elution utilizes the traditional chromatography principle of multi-equilibrium for elution of the plasmid DNA, which has a slow off-rate.
How many samples can I set up and walk away?
For maxi, mega, and gigaprep, it is possible to setup 4, 4, and 2 samples, respectively, for walk-away purification.
How many Maxi, Mega or Giga preps can I process in a day?
For Maxi, Mega and Giga preps, it is possible to purify 12, 8, and 4 samples in a day, respectively.
Can I run the AutoPlasmid MMG overnight?
Yes, it is possible to setup the MMG and complete the run unsupervised.
How can I increase my yields?
The flexibility and robustness of the AutoPlasmid MMG and kits mean that ideal mass of plasmid can be achieved. Generally, if more plasmid is desired, the user can grow larger cultures.
What are the expected yields?
The maximum yields from a Maxi, Mega and Giga prep are 1.5 mg, 5 mg, and 10 mg, respectively. The typical yields from a Maxi, Mega and Giga prep are 750 µg, 3.5 mg, and 7 mg, respectively.
What are the expected concentrations from a purification?
The concentration of plasmid DNA is dependent on the elution volume used during the process. For Maxi, Mega, and Gigapreps, concentrations can reach 1mg/mL
What are the A260/A280 and A230/A260 ratios?
The expected A260/A280 and A230/A260 ratios are 1.8-1.9 and 2.0-2.1, respectively.
What is the resulting concentration of endotoxin in the purified sample?
Endotoxin levels are less than 10 EU/µg
Are the plasmids transfection grade?
Yes, plasmids purified by PhyTip columns are transfection grade
Is it necessary to perform ethanol precipitation at the end?
No, ethanol is not necessary. Endotoxin levels are within transfection range and concentrations are high enough for downstream use.
Are the purified samples free of ethanol?
Yes, samples are free of ethanol and measured to be less than 3%.
Are the purified plasmids sterile?
Yes, samples are sterile. However, the AutoPlasmid MMG is an open instrument. It is recommended to process the samples through a post-purification kit which consists of a 0.2 micron syringe filter.
PhyNexus, the long time leader in automated solutions for protein purification and sample prep, introduces the AutoPlasmid MMG. The AutoPlasmid MMG is the industry’s first and only fully-automated plasmid purification instrument capable of Maxi-, Mega- or Gigaprep scale plasmid purification.
Transfection Quality Plasmids
It only takes about 5-10 minutes hands-on time to set up the instrument, pour in the prepared sample, hit start and walk away. Come back at the end of the run and your transfection-quality plasmids are waiting for you—no vacuums, no centrifugation.
Mega, Maxi or Gigaprep Scale
Sometimes you need more, sometimes less, so the AutoPlasmid MMG can do whatever you need—not the other way around. Set up and run up to four Maxi or Megapreps or two Gigapreps a run.
Increased Productivity
Lab professionals can be deployed to more productive, more valuable tasks
Significant Cost Benefits
Not just more productivity from staff, kit pricing comparable with manual kits and vastly cheaper than outsourcing
Save Time & Keep Control
Purified plasmids in hours, not days or weeks from outsourcing, and you keep control throughout the process
Better Sample Quality
Improved process repeatability, precision and reliability
Easy to Use
About 5-10 minutes set up, a no clean up with disposable kits
To address the trend for advanced biological research, people require large amounts of transient transfection quality plasmid DNA. The AutoPlasmid MMG is only automation instrument designed specifically to meet this market opportunity. A larger 20 mL column with 1-5 mL beds are used for higher capacity for Maxi, Mega, and Giga scale plasmid preps, all in one instrument.
Buffer kits and system software are completely adaptable to the sample and the scale desired.
The MMG provides a pushbutton, walkaway solution, including transient transfection quality plasmids.
The instrument is based on Dual Flow Chromatography, a proprietary technology of PhyNexus.
With Dual Flow Chromatography fluid flows back and forth through the pipette tip column. Interactions of plasmids with the column are pushed to completion. Through this fluid motion, columns are completely loaded allowing high recovery of the plasmids.
The AutoPlasmid MMG provides consistency and reliability due to the ability to purify in the presence of particulate. The process is rugged – there are no restrictions on the type of plasmid sample. 1-4 samples can be processed per run. Depending on scale, 3-12 preps can be performed every day. Recoveries of the purified plasmids are into 4 flip top vials with a bio compatible buffer. The plasmids are transient transfection quality and can be used for any desired application.
Prepping the instrument for any scale level is done in minutes. The system is rugged. High quality plasmid can be recovered from virtually any volume or type of sample. And when you’re done, samples trays, buffers, and columns are thrown away. No cleaning is needed to prep the instrument for the next run.
AutoPlasmid MMG Specifications
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Connections |
2 USB 1 Ethernet 1 DC Power Input |
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Instrument dimensions |
H 850 mm x W 540 mm x D740 mm (H 33.5 x W 21.3 x D 29.1 in) |
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Operating temperature |
15–32°C (59–90°F) |
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Power |
100–240 V AC; from external power supply to the instrument : 12 VDC and 24 VDC nominal |
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Weight |
41 kg (90 lbs) |
