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DNA & RNA Purification

GenElute™ HP Plasmid Kits

Description

The GenElute™ HP Plasmid Midiprep and Maxiprep Kits offer purification of high-quality plasmid DNA in less than 30 minutes (Figure 1). GenElute HP Plasmid Kits include a filter syringe for lysate clearing, binding columns that can be used with a vacuum or spin purification format, and does not require alcohol precipitation.

  • Fast – high-quality plasmid DNA in 30 minutes
  • Convenient – vacuum or spin column format
  • Cost effective – more value for the money

Procedure

An overnight recombinant E. coli culture is harvested by centrifugation and subjected to a modified alkaline-SDS lysis procedure. The lysate is cleared with a filter syringe and DNA is bound to the silica-based membrane. The remaining contaminates are removed by a wash step. Finally, the bound DNA is eluted in buffer or water.

Application

High-quality DNA is ready for immediate use for the most demanding applications, including transfection (Figure 3), restriction digestion (Figure 4), ligation, automated sequencing (Figure 5).

Beneficial Features


Culture Volume 1-5 ml (Mini), 50 ml (Midi) and 150 ml (Maxi) bacterial culture
Yield Up to 25 µg (Mini), 350 µg (Midi) and 1.2 mg (Maxi) yield of high-copy plasmid DNA (Figure 2)
Speed From harvested bacterial culture to pure plasmid DNA in 30 minutes or less
Flexible Offers the flexibility of a vacuum or spin format; does not rely on gravity flow or ultracentrifugation
Economic Lower cost per prep plus higher yields per prep equals double savings

Order Information

GenElute HP Plasmid DNA Kits

Product Code Product Name Purifications per kit Technical Manual .pdf Short Protocol .pdf
NA0150S GenElute HP Plasmid Miniprep Kit 10 X -
NA0150 GenElute HP Plasmid Miniprep Kit 70 X -
NA0160 GenElute HP Plasmid Miniprep Kit 350 X -
NA0200S GenElute HP Plasmid Midiprep Kit 4 X X
NA0200 GenElute HP Plasmid Midiprep Kit 25 X X
NA0300S GenElute HP Plasmid Maxiprep Kit 4 X X
NA0300 GenElute HP Plasmid Maxiprep Kit 10 X X
NA0310 GenElute HP Plasmid Maxiprep Kit 25 X X

Sample Data

Preparation Time and Plasmid DNA Yield for different Maxiprep Kits

Graph for Preparation Time


Figure 1. Comparison of time required per isolation using different Maxiprep plasmid isolation systems. Plasmid DNA was isolated following each manufacturers recommended protocol. Each system was tested multiple times and the average preparation time is shown here.

 

Graph for Plasmid DNA Yield


Figure 2. Comparison of plasmid DNA yield using different Maxiprep plasimd isolation systems. Plasmid DNA was isolated following each manufacturers recommended protocol. All samples were done in duplicate.

High-quality DNA for transfections

Graph for transfection of cells


Figure 3. Comparison of transfection efficiencies of plasmids prepared with different plasmid isolation methods. Data shows an average of 3 replicates for each named isolation method. All transfections were in CHO-K1 cells. The OD measurements were taken at 420 nm recording the units of β-gal/plate. The cells were grown to 60-70% confluency and transfected using 3 µg of plasmid pSPORT-CMV/15 µg of Escort IV (Product No. L3287). β-Gal activity was determined 54 hours post-transfection.

 

Restriction Enzyme Results


Figure 4. Restriction digestion of plasmid DNA isolated using the GenElute HP Plasmid Maxiprep Kits and a comparable kit from Supplier Q. The plasmid DNA (800 ng) was digested with 10 units of Hind III at 37 °C for 4 hours. Each plasmid sample represents 100 ng of undigested DNA (left) and digested DNA (right) loaded in each pair of lanes. Samples were loaded on a 1% TBE agarose gel. The marker (M) used was 1 kb DNA Ladder (Product No. D0428).

 

High-quality DNA for Automated Sequencing

Figure 5. Electropherogram revealing >600 bases of sequence from pCMV-SPORT-β-gal purified from GenElute HP Plasmid Midiprep Kit. Cycle Sequencing was performed using 500 ng template, a T7 sequencing primer and ABI BigDye™ terminator chemistry. Sequencing reactions were resolved on an ABI Prism XL instrument with a 48 cm gel cassette containing 4.5% AutoPAGE Plus acrylamide at 2.4 kV, 48°C for 7 hours.

Click Image to View Sequence in its Entirety

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