Introduction
HiPure Plasmid EF Maxi Kit B functions as the primary low-endotoxin column reference model within the Magen plasmid DNA extraction portfolio. Built as an upgraded version of the earlier EF maxi workflow, it retains the broad vector compatibility and low-endotoxin preparation logic of the original system while further improving endotoxin removal efficiency, workflow flexibility and RNA control. The product is positioned for medium- to low-copy plasmids, routine cell transfection, animal injection and other downstream applications requiring cleaner plasmid preparation at larger scale.
Compared with the earlier EF maxi format, this version introduces an upgraded endotoxin-removal workflow, a higher-capacity RC7 maxi column and a more flexible operating structure that supports either centrifugation or vacuum-based handling. Within the Magen plasmid system, HiPure Plasmid EF Maxi Kit B serves as the primary low-endotoxin column route. Laboratories working below this scale may refer to HiPure Plasmid EF Mini Kit (P1154) or HiPure Plasmid EF Midi Kit (P1231), while HiPure Plasmid EF Maxi Kit (P1156) remains available as the earlier support configuration within the same EF branch.
Details
Extraction Principle
Bacterial cells are lysed under alkaline conditions and neutralized to produce a clarified lysate suitable for endotoxin-oriented purification. The lysate is then treated with a dedicated endotoxin-removal step before column binding. For more sensitive downstream applications, the workflow also provides an optional enhanced cleanup process prior to loading. After binding conditions are adjusted with alcohol, plasmid DNA is captured on the maxi column, washed and eluted to recover low-endotoxin plasmid DNA.
Engineering Characteristics
Upgraded low-endotoxin purification workflow
This version builds on the earlier EF maxi route but further improves endotoxin removal and RNA control, making it better suited to more demanding downstream applications
RC7 maxi column with higher binding capacity
The upgraded maxi column is specified with binding yield up to 1500 μg, which is better aligned with larger-output plasmid preparation than the earlier EF maxi format.
Two operating routes for larger-scale handling
The workflow supports both centrifugation and vacuum-based processing, making it easier to integrate into different laboratory setups for routine larger-volume preparation.
Improved endotoxin and RNA control
The upgraded workflow refines the Triton X-114-based endotoxin removal step and adds a wash stage aimed at further reducing endotoxin and RNA carryover in the final plasmid DNA.
Large-format elution control
The protocol gives explicit elution guidance based on centrifuge type and plasmid size. Plasmids above 10 kb benefit from preheated elution buffer, and the recommended elution volume changes depending on whether angle or horizontal centrifugation is used. A second elution step is also supported to improve recovery.
Simplified drying after washing
Internal workflow verification showed that column drying can be streamlined after washing, helping reduce residual ethanol while simplifying routine handling.
Technical Validation
Internal validation compared centrifugation and vacuum filtration workflows across 50, 100 and 200 mL LB cultures containing pcDNA3.1 plasmid. Both processing formats showed comparable purity and recovery, with A260/280 typically around 1.89–1.95 and A260/230 around 2.02–2.30. At 200 mL culture input, total plasmid yield reached approximately 1.3–1.5 mg, supporting the product’s role as the primary large-scale low-endotoxin column workflow in the current plasmid system.
The validation results also showed that both operation modes produced broadly similar output across the tested input range, which supports the practical flexibility of this upgraded EF maxi format in routine laboratory use.
Kit Contents
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Contents
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P115602B
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P115603B
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Purification Times
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10 Preps
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50 Preps
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RNase A
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30 mg
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150 mg
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Buffer P1
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100 ml
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500 ml
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Buffer P2
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90 ml
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450 ml
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Buffer NS3
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90 ml
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450 ml
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Buffer PW1
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60 ml
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270 ml
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Buffer PW2*
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20 ml
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100 ml
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Elution Buffer
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20 ml
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120 ml
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Buffer ER2
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30 ml
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150 ml
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HiPure DNA Maxi Columns RC7
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10
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50
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Lysate Clear Midi Syringe
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10
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50
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50 ml Collection Tube
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20
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100
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Storage and Stability
The kit components can be stored dry at room temperature (15–25°C) and are stable for at least 18 months under these conditions. If any precipitates form in the buffers,warm at 37°C to dissolve. After addition of RNase A，Buffer P1 is stable for 6 months when stored at 2–8°C.
